[go: up one dir, main page]

WO2025221360A1 - Systems and methods for generating lipid nanoparticles - Google Patents

Systems and methods for generating lipid nanoparticles

Info

Publication number
WO2025221360A1
WO2025221360A1 PCT/US2025/016069 US2025016069W WO2025221360A1 WO 2025221360 A1 WO2025221360 A1 WO 2025221360A1 US 2025016069 W US2025016069 W US 2025016069W WO 2025221360 A1 WO2025221360 A1 WO 2025221360A1
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
static mixer
lipid
inlet
conduit
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/US2025/016069
Other languages
French (fr)
Inventor
Neda MAHMOUDI
Jacob Kenneth RUEBEN
Sakya Sing Mohapatra
Vladimir Grigor'evich KHARITONOV
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Recode Therapeutics Inc
Original Assignee
Recode Therapeutics Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Recode Therapeutics Inc filed Critical Recode Therapeutics Inc
Publication of WO2025221360A1 publication Critical patent/WO2025221360A1/en
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
    • B01F23/40—Mixing liquids with liquids; Emulsifying
    • B01F23/41—Emulsifying
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
    • B01F25/40—Static mixers
    • B01F25/42—Static mixers in which the mixing is affected by moving the components jointly in changing directions, e.g. in tubes provided with baffles or obstructions
    • B01F25/43—Mixing tubes, e.g. wherein the material is moved in a radial or partly reversed direction
    • B01F25/431—Straight mixing tubes with baffles or obstructions that do not cause substantial pressure drop; Baffles therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/30—Micromixers
    • B01F33/301—Micromixers using specific means for arranging the streams to be mixed, e.g. channel geometries or dispositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/5123—Organic compounds, e.g. fats, sugars
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
    • B01F35/20—Measuring; Control or regulation
    • B01F35/21—Measuring
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/02—Making microcapsules or microballoons
    • B01J13/04—Making microcapsules or microballoons by physical processes, e.g. drying, spraying

Definitions

  • Atty Dkt No.: 58530-742601 SYSTEMS AND METHODS FOR GENERATING LIPID NANOPARTICLES CROSS-REFERENCE
  • This application claims benefit of U.S. Provisional Application No. 63/554,054, filed February 15, 2024, and U.S. Provisional Application No.63/737,069, filed December 20, 2024, each of which is incorporated by reference herein in its entirety.
  • Therapeutic nucleic acids such as those that enable gene silencing, expression and editing possess great potential for use as genetic medicines.
  • Lipid nanoparticles may be used for delivery of nucleic acids for treatment or prevention of various diseases and disorders.
  • the system comprises a conduit, a static mixer, wherein a nucleic acid and one or more lipid components are mixed to generate a mixture fluid; and a high performance liquid (HPLC) pump.
  • HPLC high performance liquid
  • a system for generating a lipid nanoparticle the system comprising a conduit and a static mixer, wherein a nucleic acid and one or more lipid components are mixed to generate a mixture fluid, wherein when the nucleic acid and the one or more lipid components are joined in the static mixer, the system reaches a pressure of no greater than 8 pounds per square inch gauge (psig).
  • the conduit comprises a first inlet configured to introduce the nucleic acid. In some embodiments, the conduit comprises a second inlet configured to introduce one or more lipid components of the lipid nanoparticle. In some embodiments, the conduit comprises an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet. In some embodiments, the conduit comprises a third inlet configured to introduce the nucleic acid or one or more lipid components of the lipid nanoparticle. In some embodiments, the conduit comprises a fourth inlet configured to introduce the nucleic acid or the one or more lipid components of the lipid nanoparticle.
  • the static mixer comprises a plurality of elements, wherein each element of the static mixer makes a 270° twist every 1/8 inch.
  • the system comprises a channel housing the static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the channel is square shaped and makes a -180° twist every 1/8 inch.
  • the conduit is tee-shaped.
  • the conduit is Y-shaped.
  • the conduit further comprises a cut zip tie.
  • the conduit is L-shaped.
  • the static mixer is less than about 40 millimeters (mm) in length.
  • the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index of the mixture fluid is about 0.1 to about 0.2. In some embodiments, the polydispersity index of the mixture fluid is no greater than about 0.1.
  • a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the diameter of the lipid nanoparticle is no greater than about 100 nanometers (nm). In some embodiments, the diameter of the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the diameter of the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit.
  • a system for generating a lipid nanoparticle comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is no greater than about 0.25.
  • the conduit is tee-shaped.
  • the Atty Dkt No.: 58530-742601 conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 millimeters (mm) in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the polydispersity index is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nanometers (nm). In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm.
  • a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements.
  • the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit comprising: a first inlet; a second inlet; an outlet; and a static mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more Atty Dkt No.: 58530-742601 lipid components, thereby generating a mixture fluid, wherein a polydispersity index of the mixture fluid is no greater than about 0.25.
  • the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the polydispersity index is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm.
  • a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements.
  • the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • a system for generating a lipid nanoparticle comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second Atty Dkt No.: 58530-742601 inlet configured to introduce one or more lipid components of the lipid nanoparticle; a junction site wherein the first inlet and the second inlet intersects; and a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site.
  • the conduit is tee-shaped.
  • the conduit is Y-shaped.
  • the conduit further comprises a cut zip tie. In some embodiments, the conduit is L- shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm.
  • the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements.
  • the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance Atty Dkt No.: 58530-742601 liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC liquid chromatography
  • a method of generating a lipid nanoparticle comprising: providing: a conduit comprising: a first inlet; a second inlet; a junction site downstream of the first inlet and the second inlet; and a static mixer, wherein the static mixer is disposed at a distal end of the junction site; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the junction site; and using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site.
  • the conduit is tee-shaped. In some embodiments, the conduit is Y- shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm.
  • the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • the Atty Dkt No.: 58530-742601 static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • a system for generating a lipid nanoparticle comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a Reynolds number of the mixture fluid is no greater than about 1000.
  • the conduit is tee-shaped.
  • the conduit is Y-shaped.
  • the conduit further comprises a cut zip tie.
  • the conduit is L-shaped.
  • the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2.
  • the polydispersity index is no greater than about 0.1.
  • the Reynolds number is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000.
  • the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • the nucleic acid and the one or more Atty Dkt No.: 58530-742601 lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements.
  • the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • a method of generating a lipid nanoparticle comprising providing a conduit and a static mixer, providing a nucleic acid and one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate, introducing the nucleic acid to the first inlet, introducing one or more lipid components of the lipid nanoparticle to the second inlet, joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid comprising the lipid nanoparticle.
  • HPLC high performance liquid chromatography
  • the conduit comprises a first inlet. In some embodiments, the conduit comprises a second inlet. In some embodiments, the conduit comprises a third inlet. In some embodiments, the conduit comprises a fourth inlet. In some embodiments, the conduit comprises an outlet. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer comprises a plurality of elements, wherein each element of the static mixer makes a 270° twist every 1/8 inch. In some embodiments, the static mixer is less than about 40 mm in length.
  • the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index of the mixture fluid is about 0.1 to about 0.2. In some embodiments, the polydispersity index of the mixture fluid is no greater than about 0.1.
  • the Reynolds number is Atty Dkt No.: 58530-742601 about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the diameter of the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the diameter of the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the diameter of the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit.
  • aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit comprising: a first inlet; a second inlet; an outlet; and a static mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid, wherein a Reynolds number of the mixture fluid is no greater than about 1000.
  • the conduit is tee-shaped.
  • the conduit is Y-shaped.
  • the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2.
  • the polydispersity index is no greater than about 0.1.
  • the Reynolds number is Atty Dkt No.: 58530-742601 about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000.
  • the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit.
  • the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • a system for generating a lipid nanoparticle comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min.
  • the conduit is tee-shaped.
  • the conduit is Y-shaped.
  • the conduit further comprises a cut zip tie.
  • the conduit is L-shaped.
  • the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch. In some embodiments, the inner diameter is about 1 ⁇ 4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the Atty Dkt No.: 58530-742601 mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm.
  • the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit.
  • the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit comprising: a first inlet; a second inlet; an outlet; and a static mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min.
  • the conduit is tee-shaped.
  • the conduit is Y-shaped.
  • the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of Atty Dkt No.: 58530-742601 about 1/16 inch to about 1 ⁇ 2 inch. In some embodiments, the inner diameter is about 1 ⁇ 4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm.
  • the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements.
  • the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • a system for generating a lipid nanoparticle comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the lipid nanoparticle is no greater than about 150 nm.
  • the conduit is tee- shaped.
  • the conduit is Y-shaped.
  • the conduit further comprises a cut zip tie.
  • the conduit is L-shaped.
  • the static mixer is less than about 40 mm in length.
  • the Atty Dkt No.: 58530-742601 static mixer is less than about 10 mm in length.
  • the static mixer is less than about 5 mm in length.
  • the static mixer is about 4.8 mm in length.
  • the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1 ⁇ 4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm.
  • the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements.
  • the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • a method of generating a lipid nanoparticle comprising: providing: a conduit comprising: a first inlet; a second inlet; an outlet; and a static mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the lipid nanoparticle is no greater than about 150 nm.
  • the conduit is tee-shaped.
  • the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L- shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch. In some embodiments, the inner diameter is about 1 ⁇ 4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm.
  • the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements.
  • the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • a system for generating a lipid nanoparticle comprising: a conduit, comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, Atty Dkt No.: 58530-742601 wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer configured to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • the conduit is tee-shaped.
  • the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch. In some embodiments, the inner diameter is about 1 ⁇ 4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm.
  • the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements.
  • the static mixer is downstream of the conduit.
  • the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit, comprising: a first inlet; a second inlet; an outlet; and a static Atty Dkt No.: 58530-742601 mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch. In some embodiments, the inner diameter is about 1 ⁇ 4 inch. In some embodiments, the inner diameter is about 1/8 inch.
  • the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm.
  • the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements.
  • the static mixer is downstream of the conduit.
  • the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
  • HPLC high performance liquid chromatography
  • Another aspect of the present disclosure provides a non-transitory computer readable medium comprising machine executable code that, upon execution by one or more computer processors, implements any of the methods above or elsewhere herein.
  • Another aspect of the present disclosure provides a system comprising one or more computer processors and computer memory coupled thereto.
  • the computer memory comprises machine executable code that, upon execution by the one or more computer processors, implements any of the methods above or elsewhere herein.
  • a system for generating a lipid nanoparticle comprising (i) a first inlet configured to introduce a nucleic acid, (ii) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle, and (iii) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein when a nucleic acid and one or more lipid components are mixed in the static mixer, the system reaches a pressure of no greater than 8 psig.
  • the static mixer is less than about 40 mm in length.
  • the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch. In some embodiments, the inner diameter is about 1 ⁇ 4 inch. In some embodiments, the inner diameter is about 1/8 inch. [0021] In some embodiments, the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1.
  • a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. [0023] In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. [0024] In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Atty Dkt No.: 58530-742601 [0025] In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. [0026] In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements.
  • the static mixer comprises 24 or more elements.
  • the method comprises diluting the lipid nanoparticle.
  • a particle size of the lipid nanoparticle is within 10% of the particle size of the lipid nanoparticle before dilution.
  • a method of generating a lipid nanoparticle comprising: (a) providing a system provided herein, (b) introducing a nucleic acid to the first inlet, (c) introducing one or more lipid components of the lipid nanoparticle to the second outlet, (d) joining the nucleic acid and the one or more lipid components at the outlet, and I using the static mixer to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle, wherein a particle size of the lipid nanoparticle is selected by modifying the Reynolds number.
  • the particle size is increased by decreasing the Reynolds number. In some embodiments, the particle size is decreased by increasing the Reynolds number.
  • FIGs. 1A-I show schematics of the nine tested static mixing setups.
  • FIG. 1A and FIG. 1B show systems A and B, both using a Koflo 24-element 1/8” ID disposable static mixer with a polypropylene tube, with A using a tee and B using a Y for joining inlets prior to mixing.
  • FIG. 1C shows system C using a Koflo 12-element 1/4” OD mixing element 1/4” ID platinum cured silicone tubing.
  • FIG. 1D shows system D uses the same mixer and Y as B, but with the mixer pushed partway into the Y.
  • FIG. 1E shows system E which uses the same mixer and Y as B, only with some inlet separation provided by a cut zip tie.
  • FIG. 1F shows system F which is the same as B, but with only 12 mixing elements.
  • FIG.1G shows system G which is a custom mixer with three 12-element 1/8” mixer together in a 3/16” ID silicone tube.
  • FIG.1H shows system H is a custom mixer with 16 mixing elements, some of which are pushed into a tee such that the lipid and buffer are joined during mixing.
  • FIG. 1I shows system I is a custom mixer with 12 mixing elements, and with the mixer pushed into the lipid inlet of a Y junction.
  • FIG. 2 shows images of food dye tests for systems A-C at Reynolds numbers of 25, 50, and 100.
  • FIGs. 3A-B show particle diameter plotted against Reynolds number for system B run at 1:2 FRR (FIG.3A) and polydispersity index (PDI) for the same samples shown in FIG.3A (FIG.3B).
  • FIGs. 4A-F shows schematics of the six static mixing setups (systems J-N). All systems were tested at Re of 1000.
  • FIG.4A shows system J which is essentially system H (FIG.1H) but with the inlet streams switched and a higher total flow rate.
  • FIG.4B shows system K which is essentially system H (FIG.1H) but with a higher total flow rate.
  • FIG.4C shows system L which is System J but with a 1:3 FRR and no in-line dilution.
  • FIG.4D shows system M which is system J but with a 1:5 FRR and no in-line dilution.
  • FIG.4E shows System N which is System J Atty Dkt No.: 58530-742601 but with a shorter connection between the two tees and half as many mixing elements.
  • FIG.4F shows System O has similar design as Systems J-M but without mixing elements.
  • FIGs. 5A-B show bar graphs of post-dilution particle diameter (FIG.
  • FIGs. 6A-B show plots of post-dilution and 15 minutes post-mix particle diameter (FIG. 6A) and PDI (FIG.6B).
  • FIGs. 7A-C shows bar graphs of post-storage particle diameter (FIG. 7A), post-storage PDI (FIG.7B), and post-storage encapsulation efficiency (FIG.7C) for each of the five tested compositions, also showing a static mixing sample compared to a previously formulated cross- tee sample.
  • FIGs. 8A-B show a splitter insert designed to split flow prior to mixing elements, ensuring consistent mixing.
  • FIGs. 9A-C show a mixer (FIGs. 9A, C) with a length of 1 inch (25.4 mm) and a diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.9B), each 1/8 inches (3.175 mm) in length that make a 180° twist. The cross-section thickness if about 0.0394 inches (1mm).
  • FIGs. 10A-B show a mixer (FIG. 10A) and mixing simulation (FIG.
  • FIGs. 11A-C show results from two different viscosity models, k-omega with SST (FIG. 11A) and transition-SST (FIG.11B), which is more rigorous for transition-range Reynolds numbers. Both models appeared similar with respect to mixing estimation, as shown by cross- sections (FIGs.11A-B) and an extrapolated plot of standard deviations of ethanol mass fraction at different cross-sections through the mixer (FIG.11C). [0044] FIGs.
  • FIGs. 12A-C show results from two different viscosity models, k-omega with SST (FIG. 12A) and transition-SST (FIG.12B). When comparing shear stress, differences between model outputs are more evident (FIG.12C). Across the board, transition-SST model outputs about 40% higher shear stress than k-omega model.
  • FIGs. 13A-B show comparison of mixing by outlet.
  • FIG. 13A shows the mass fraction of ethanol for outflow.
  • FIG.13B shows mass fraction of ethanol for the pressure-outlet.
  • FIGs. 14A-B show comparison of shear stress between outlets.
  • FIG. 14A shows shear stress for the outflow
  • FIG.14B show shear stress for the pressure-outlet.
  • FIGs. 15A-C show a mixer with 8 mixing elements.
  • FIG. 15A shows a 3D model of the mixer with eight mixing elements with dimensions, each making a 180° twist.
  • FIG.15B shows an example mixing conduit 3D model made with mixing elements rendered in FIG.15A.
  • FIG. 15C shows a 3D model of fluid volume/path of mixing conduit shown in FIG.15B.
  • FIGs. 16A-B show a 3D rendering of a 0.5 mm bore size cross mixer fluid volume.
  • FIG. 16A shows an isometric view.
  • FIG.16B shows a top view.
  • FIG.17A-C show results comparing an 8-element static mixer to a 0.5 mm cross mixer.
  • FIG.17A shows comparison of standard deviation of ethanol mass fraction between the 8- element static mixer and the 0.5 mm cross mixer. Time point of zero indicates first time of mixing between ethanol and water streams, and each data point represents the standard deviation across a cross-section perpendicular to flow direction.
  • FIG.17B shows comparison of average shear stress between the 8-element static mixer and the 0.5 mm cross mixer. Each data point represents the area-weighted average shear stress across a cross-section perpendicular to flow direction.
  • FIG.17C shows the maximum system pressure for both the 8-element static mixer and the 0.5 mm cross mixer. [0050] FIGs.
  • FIGs. 19A-C show a mixer (FIG. 18A, C) with a length of 0.8 inches (20.32 mm) and diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.18B) each 0.1 inches (2.54 mm) in length, each element making a 180° twist.
  • the cross-section thickness is about 0.0394 inches (1 mm).
  • FIGs. 19A-C show a mixer (FIG. 19A, C) with a length of 1.2 inches (30.48 mm) and diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.19B) each 0.15 inches (3.81 mm) in length, where each element makes a 180° twist.
  • the cross-section thickness is about 0.0394 inches (1 mm).
  • FIGs. 20A-C show a 3-channel mixer (FIG. 20A, C) with a length of 1 inch (25.4 mm) and diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.20B), each element 1/8 inches in length, and each element making a 180° twist.
  • the cross-section thickness is 0.0197 inches (0.5 mm) and is Y shaped.
  • FIGs. 21A-C show a 4-channel mixer (FIG. 21A, C) with a length of 1 inch (25.4 mm) and diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.21B), each element 1/8 inches in length, and each element making a 180° twist.
  • FIGs. 22A-C show a 90° turn mixer (FIG. 22A, C) with a length of 1 ⁇ 2 inches (12.7 mm) comprising two channels and is 1/8 inches (3.175 mm) in diameter.
  • the mixer comprises 8 Atty Dkt No.: 58530-742601 elements (FIG.22B), each 1/16 inches (1.59 mm) in length and each making a 90° twist.
  • the cross-section thickness is about 1 mm.
  • FIGs. 23A-C show a 270° turn mixer (FIGs.
  • FIG. 24 shows an exemplary static mixing setup with a conduit 2401 and a mixer 2402.
  • the conduit can comprise a first inlet 2403, a second inlet 2404, an outlet 2405, and a junction site 2406.
  • the conduit can have an angle 2407 between the first inlet 2403 and the second inlet 2404.
  • the conduit can have an angle 2408 between the first inlet 2403 and the outlet 2405.
  • FIGs. 25A-C show a Koflo analogue design mixer (Design 1, No. A1), with 2 inlets and helical elements and 180° twists (FIG.25A) and a cylindrical channel that houses the elements, and comprise of 1/8 inch barbs that are 120° apart (FIG.25B, C).
  • FIGs. 26A-D show a design mixer (Design 2, No. D10), with 2 inlets and helical elements and 270° twists (FIG.26A, B) and a square shaped channel that houses the elements (FIG.26C) and comprise -180° twists (FIG.26D).
  • FIGs. 27A-D show a design mixer (Design 3, No. D7), with 4 inlets and helical elements and 270° twists (FIG.27A, B) and a square shaped channel that houses the elements (FIG.27C) and comprise -180° twists (FIG.27D).
  • FIGs. 28A-D show a design mixer (Design 5, No. D6), with 4 inlets and helical elements and 180° twists (FIG.28A, B) and a square shaped channel that houses the elements (FIG.28C) and comprise -180° twists (FIG.28D).
  • FIG. 29A, FIG. 29B, and FIG. 29C show static mixers as described herein.
  • nucleic acids that enable gene silencing, expression, and editing possess great potential for use in genetic medicines for treatment of various indications, including cancer, inherited genetic disorders, and infectious diseases.
  • Efficacious application of nucleic acids for treatment of the abovementioned classes of disorders generally require the use of viral and non-viral delivery methods to facilitate delivery of the nucleic acid to target cells.
  • Lipid nanoparticles (LNPs) represent a potentially efficacious non-viral delivery platform for nucleic acids.
  • Atty Dkt No.: 58530-742601 generating LNPs comprising nucleic acids.
  • LNP preparation often faces several challenges including particle size and dispersity control.
  • the effectiveness of LNPs can depend on their size and it can be challenging to consistently prepare particles of the size needed for various applications.
  • LNP preparation may also face the issue of low reproducibility due to the complex production processes involved. Cost and scale-up may also inhibit clinical translation of LNPs as preparation of LNPs at scale may be met with reduced homogeneity or modified properties.
  • LNPs are prepared by various methods including high pressure homogenization, microemulsion, solvent emulsification/evaporation, and ultrasonication.
  • the present disclosure provides systems and methods for generation of LNPs.
  • the LNPs may have a small particle size.
  • the LNPs may have a low polydispersity index in a solution.
  • the systems and methods provided herein may be low-cost and may be scaled up efficiently.
  • Methods of generating LNPs as described herein may comprise using a conduit with one or more inlets.
  • the conduit may have one or more outlets.
  • the conduit may have a static mixer.
  • the static mixer may have improved scalability compared to alternative setups (e.g., tee, cross, or microfluidic setups).
  • the improved scalability may be important for commercial scale manufacturing.
  • Each element may allow for production of LNPs with varying ratios of components.
  • the methods provided herein may utilize lower flow and back-pressure and may aid in achieving better control over the physicochemical properties of the resulting drug product.
  • the use of lower flow and lower back-pressure regime could provide a broader selection of pump options and a better control on LNP formation.
  • the use of static mixers provided herein may be a suitable choice for LNP generation.
  • the methods provided herein may allow for reproducible preparation of LNPs with increased control over properties such as size, monodispersity, scalability, and drug loading. Furthermore, the methods provided herein are versatile, allowing for the substitution, addition, or removal of various LNP components (e.g., allowing for 3-component LNPs, 4-component LNPs, 5-component LNPs, or the like) without the need to sacrifice control over the aforementioned properties.
  • Systems and Methods for Generating Lipid Nanoparticles [0064] The present disclosure provides systems and methods for generating lipid nanoparticles. In some embodiments, the lipid nanoparticle encapsulates a nucleic acid.
  • the nucleic acid may be, for example, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or Atty Dkt No.: 58530-742601 DNA copies of ribonucleic acid (cDNA).
  • the RNA may be, for example, messenger RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), guide RNAs (gRNAs), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), SmY RNA, spliced leader RNA (SL RNA), CRISPR RNA, long noncoding RNA (lncRNA), microRNA (miRNA), or another suitable RNA.
  • the nucleic acid may be RNA.
  • the nucleic acid may be mRNA.
  • the nucleic acid may be DNA.
  • Conduit [0065]
  • Systems as disclosed herein may comprise a conduit (e.g., as shown as 2401 in FIG. 24).
  • the conduit comprises one or more inlets (e.g., a first inlet, as shown as 2403 in FIG.24, and a second inlet, as shown as 2404 in FIG.24), an outlet, as shown as 2405 in FIG.24, and a junction site, as shown as 2406 in FIG.24.
  • the conduit comprises ten or more inlets. In some embodiments, the ten or more inlets intersect at a junction site.
  • the conduit comprises one to ten (e.g., or more) inlets. In some embodiments, the conduit comprises one to eight inlets. In some embodiments, the conduit comprises one to five inlets. In some embodiments, the conduit comprises one to three inlets. In some embodiments, the conduit comprise one inlet. In some embodiments, the conduit comprises two inlets. In some embodiments, the conduit comprises three inlets. In some embodiments, the conduit comprises four inlets. In some embodiments, the conduit comprises five inlets. In some embodiments, the conduit comprises six inlets. In some embodiments, the conduit comprises seven inlets. In some embodiments, the conduit comprises eight inlets. In some embodiments, the conduit comprises nine inlets. In some embodiments, the conduit comprises ten inlets.
  • the conduit comprises ten inlets.
  • the conduit may be tee-shaped, as shown in FIG. 1A.
  • the conduit may be Y-shaped, as shown in FIGs. 1B-1G, and FIG. 1I.
  • the Y-shaped conduit may further comprise a cut zip tie, as shown in FIG.1E.
  • the cut zip tie may provide inlet separation.
  • the cut zip tie may comprise a makeshift flow splitter.
  • the conduit may be L- shaped, as shown in FIG.1H, and FIGs.4A-4F.
  • Each of the inlets of the conduit may be used to introduce one or more components of the lipid nanoparticle.
  • each inlet is used to introduce a single component of the lipid nanoparticle. In some instances, an inlet is used to introduce at least two (e.g., 3, 4, or 5) components of the lipid nanoparticle. In some embodiments, an inlet is used to introduce a therapeutic agent (e.g., a nucleic acid) and a second (e.g., or second, third, fourth, and/or fifth) inlet is used to introduce the other components of the lipid nanoparticle.
  • a therapeutic agent e.g., a nucleic acid
  • a second inlet e.g., or second, third, fourth, and/or fifth
  • the conduit may comprise a first inlet, as shown as 2403 in FIG. 24.
  • the first inlet may be configured to introduce a nucleic acid.
  • the first inlet may be configured to introduce one or more lipid components of the lipid nanoparticle.
  • the conduit may comprise a second inlet, as shown as 2404 in FIG. 24.
  • the second inlet may be configured to introduce a nucleic acid.
  • the second inlet may be configured to introduce one or more lipid components of the lipid nanoparticle.
  • the conduit may comprise a third inlet.
  • the third inlet may be configured to introduce a nucleic acid.
  • the third inlet may be configured to introduce one or more lipid components of the lipid nanoparticle.
  • the conduit may comprise a fourth inlet.
  • the fourth inlet may be configured to introduce a nucleic acid.
  • the fourth inlet may be configured to introduce one or more lipid components of the lipid nanoparticle.
  • the conduit may comprise an outlet, as shown as 2405 in FIG. 24.
  • the conduit may comprise an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet.
  • the conduit may comprise a junction site, wherein the first inlet and the second inlet intersects, as shown as 2406 in FIG.24. In some embodiments, the nucleic acid and one or more lipid components may not contact each other prior to entering the junction site.
  • the conduit may comprise an angle, as shown as 2407 in FIG. 24, between the first inlet and the second inlet.
  • the angle between the first inlet and the second inlet may be no more than 210° (e.g., no more than 180°, 160°, 140°, 120°, 100°, 90°, 80°, 70°, 60°, 50°, 40°, 30°, 20°, or 10°). In some embodiments, the angle between the first inlet and the second inlet is at least 10° (e.g., at least 15°, 20°, 25°, 30°, 35°, 40°, 45°, 50°, 60°, 70°, 80°, or 90°).
  • the angle between the first inlet and the second inlet is from about 10° to about 90° (e.g., about 10° to about 60°, about 10° to about 45°, about 20° to about 90°, about 30° to about 90°, or about 20° to about 45°). In some embodiments, the angle between the first inlet and the second inlet is about 10°, about 15°, about 20°, about 25°, about 30°, about 35°, about 40°, about 45°, about 50°, about 55°, about 65°, about 70°, about 75°, about 80°, or about 90°. In some embodiments, the angle is less than 100°. In some embodiments, the angle is less than 90°.
  • the conduit may comprise an angle, as shown as 2408 in FIG. 24, between the first inlet and the outlet. In some embodiments, the angle between the first inlet and the outlet may be at Atty Dkt No.: 58530-742601 least 10° (e.g., at least 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, or 170°).
  • the angle between the first inlet and the outlet is at most 180° (e.g., at most 170°, 160°, 150°, 140°, 130°, 120°, 110°, 100°, 90°, 80°, 70°, 60°, 50°, or 40°). In some embodiments, the angle between the first inlet and the outlet is from about 40° to about 180° (e.g., 40° to about 160°, 60° to about 160°, 80° to about 160°, or about 100° to about 150°.
  • the angle between the first inlet and the second outlet is about 180°, about 170°, about 160°, about 150°, about 140°, about 130°, about 120°, about 110°, about 100°, about 90°, or about 80°. In some embodiments, the angle is at least 90°. In some embodiments, the angle is at least 120°. In some embodiments, the angle is about 100° to about 170°. [0077]
  • the conduit may comprise an angle, as shown as 2409 in FIG. 24, between the second inlet and the outlet.
  • the angle between the second inlet and the outlet may be at least 10° (e.g., at least 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, or 170°). In some embodiments, the angle between the second inlet and the outlet is at most 180° (e.g., at most 170°, 160°, 150°, 140°, 130°, 120°, 110°, 100°, 90°, 80°, 70°, 60°, 50°, or 40°).
  • the angle between the second inlet and the outlet is from about 40° to about 180° (e.g., 40° to about 160°, 60° to about 160°, 80° to about 160°, or about 100° to about 150°. In some embodiments, the angle between the second inlet and the second outlet is about 180°, about 170°, about 160°, about 150°, about 140°, about 130°, about 120°, about 110°, about 100°, about 90°, or about 80°. In some embodiments, the angle is at least 90°. [0078] In some embodiments, the angle between the first inlet and the outlet, and the angle between the second inlet and the outlet are the same.
  • the angle between the first inlet and the outlet, and the second inlet and the outlet are different.
  • the angle between an inlet (e.g., such as a first inlet, second inlet, third inlet, fourth inlet, fifth inlet, etc.) and the outlet may be at least 10° (e.g., at least 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, or 170°).
  • the angle between an inlet (e.g., such as a first inlet, second inlet, third inlet, fourth inlet, fifth inlet, etc.) and the outlet is at most 180° (e.g., at most 170°, 160°, 150°, 140°, 130°, 120°, 110°, 100°, 90°, 80°, 70°, 60°, 50°, or 40°).
  • the angle between an inlet (e.g., such as a first inlet, second inlet, third inlet, fourth inlet, fifth inlet, etc.) and the outlet is from about 40° to about 180° (e.g., 40° to about 160°, 60° to about 160°, 80° to about 160°, or about 100° to about 150°.
  • the angle between an inlet (e.g., such as a first inlet, second inlet, third inlet, fourth inlet, fifth inlet, etc.) and the second outlet is about Atty Dkt No.: 58530-742601 180°, about 170°, about 160°, about 150°, about 140°, about 130°, about 120°, about 110°, about 100°, about 90°, or about 80°.
  • a flow rate of the lipid component into any of the inlets herein may be no greater than about 500 mL/min, 300 mL/min, 200 mL/min, 150 mL/min, 100 mL/min, 50 mL/min, 40 mL/min, 30 mL/min, 20 mL/min, 10 mL/min, or 5 mL/min.
  • a flow rate of the lipid component into any of the inlets herein is at least 5 mL/min, 10 mL/min, 20 mL/min, 30 mL/min, 40 mL/min, 50 mL/min, 60 mL/min, or 70 mL/min.
  • a flow rate of the lipid component any of the inlets herein is from about 5 mL/min to about 100 mL/min, about 5 mL/min to about 80 mL/min, about 10 mL/min to about 80 mL/min, about 20 mL/min to about 80 mL/min, about 30 mL/min to about 80 mL/min, about 40 mL/min to about 80 mL/min, or about 50 mL/min to about 80 mL/min.
  • a flow rate of the lipid component into any of the inlets herein is from about 50 mL/min to about 80 mL/min. Increased flow rates may be useful to allow adoption into larger scale production.
  • a flow rate of the nucleic acid into the first inlet may be no greater than about 1200 milliliters per minute (mL/min), 1100 mL/min, 1000 mL/min, 900 mL/min, 800 mL/min, 700 mL/min,500 mL/min, 300 mL/min, 200 mL/min, 150 mL/min, 100 mL/min, 50 mL/min, 40 mL/min, 30 mL/min, 20 mL/min, 10 mL/min, or 5 mL/min.
  • mL/min milliliters per minute
  • a flow rate of the nucleic acid into the first inlet may be at least 1 mL/min, 5 mL/min, 10 mL/min, 20 mL/min, 30 mL/min, 40 mL/min, 50 mL/min, or 100 mL/min.
  • a flow rate of the nucleic acid into the first inlet may be from about 5 mL/min to about 300 mL/min, from about 5 mL/min to about 200 mL/min, from about 20 mL/min to about 200 mL/min, from about 40 mL/min to about 200 mL/min, from about 50 mL/min to about 200 mL/min, from about 100 mL/min to about 200 mL/min, or from about 120 mL/min to about 190 mL/min.
  • the flow rate of the nucleic acid into the first inlet is about 10 mL/min, such as described in Table 5.
  • the flow rate of the nucleic acid into the first inlet is about 150 mL/min to about 160 mL/min, such as described in Table 5. In some embodiments, the flow rate of the nucleic acid into the first inlet is about 150 mL/min, such a described in Table 5 or Table 7. In some embodiments, the flow rate of the nucleic acid into the first inlet is from about 200 mL/min to about 700 mL/min.
  • a flow rate of the lipid component into the second inlet may be no greater than about 1200 mL/min, 1100 mL/min, 1000 mL/min, 900 mL/min, 800 mL/min, 700 mL/min, 500 mL/min, 300 mL/min, 200 mL/min, 150 mL/min, 100 mL/min, 50 mL/min, 40 mL/min, 30 mL/min, 20 mL/min, 10 mL/min, or 5 mL/min.
  • a flow rate of the lipid component into the second inlet is at least 5 mL/min, 10 mL/min, 20 mL/min, 30 mL/min, 40 mL/min, 50 mL/min, 60 mL/min, 70 mL/min, 100 mL/min, 250 mL/min, 300 mL/min, 400 mL/min, 500 mL/min, 600 mL/min, 700 mL/min, or 800 mL/min.
  • a flow rate of the lipid component into the second inlet is from about 5 mL/min to about 1200 mL/min, about 100 mL/min to about 1000 mL/min, about 5 mL/min to about 100 mL/min, about 5 mL/min to about 80 mL/min, about 10 mL/min to about 80 mL/min, about 20 mL/min to about 80 mL/min, about 30 mL/min to about 80 mL/min, about 40 mL/min to about 80 mL/min, or about 50 mL/min to about 80 mL/min.
  • a flow rate of the lipid component into the second inlet is from about 50 mL/min to about 80 mL/min. In some embodiments, a flow rate of the lipid component into the second inlet is about 75 mL/min, such as described in Table 5 and Table 7. In some embodiments, a flow rate of the lipid component into the second inlet is about 55 mL/min, such as described in Table 5. In some embodiments, a flow rate of the lipid component into the second inlet is about 5, such as described in Table 5. In some embodiments, a flow rate of the lipid component into the second inlet is from about 50 mL/min to about 300 mL/min.
  • a ratio between a flow rate of the lipid component into any two of the inlets provided herein is from about 1:1 to about 1:10, about 1:1 to about 1:8, about 1:1 to about 1:7, about 1:1 to about 1:6, about 1:1 to about 1:3 or about 1:1 to about 1:4.
  • a ratio between a flow rate of the lipid component into any two of the inlets provided herein is at least about 1:1, 1:1.25, 1:1.5, 1:1.75, 1:2, 1:2.5, 1:3, or 1:4.
  • a ratio between a flow rate of the lipid component into any two of the inlets provided herein is at most about 1:10, 1:8, 1:6, 1:5, 1:4, 1:3, or 1:2.
  • a ratio between a flow rate of the lipid component into any two of the inlets provided herein may be about 1:2, about 1:3, about 1:5, about 1:8, or about 1:10.
  • a ratio between a flow rate of the lipid component into the second inlet and a flow rate of the nucleic acid into the first inlet is from about 1:1 to about 1:10, about 1:1 to about 1:8, about 1:1 to about 1:7, about 1:1 to about 1:6, about 1:1 to about 1:3 or about Atty Dkt No.: 58530-742601 1:1 to about 1:4.
  • a ratio between a flow rate of the lipid component into the second inlet and a flow rate of the nucleic acid into the first inlet is at least about 1:1, 1:1.25, 1:1.5, 1:1.75, 1:2, 1:2.5, 1:3, or 1:4.
  • a ratio between a flow rate of the lipid component into the second inlet and a flow rate of the nucleic acid into the first inlet is at most about 1:10, 1:8, 1:6, 1:5, 1:4, 1:3, or 1:2. In some embodiments, a ratio between a flow rate of the lipid component into the second inlet and a flow rate of the nucleic acid into the first inlet may be about 1:2, about 1:3, about 1:5, about 1:8, or about 1:10. Inlet to Introduce Nucleic Acid [0085]
  • the first inlet may be configured to introduce a therapeutic agent, such as a nucleic acid. In some embodiments, the first inlet may be configured to introduce a nucleic acid.
  • the second inlet may be configured to introduce a therapeutic agent, such as a nucleic acid. In some embodiments, the second inlet may be configured to introduce a nucleic acid. In some embodiments, the third inlet may be configured to introduce a therapeutic agent, such as a nucleic acid. In some embodiments, the third inlet may be configured to introduce a nucleic acid. In some embodiments, the fourth inlet may be configured to introduce a therapeutic agent, such as a nucleic acid. In some embodiments, the fourth inlet may be configured to introduce a nucleic acid.
  • the nucleic acid may comprise a small interfering ribonucleic acid (siRNA), a short hairpin RNA (shRNA), a micro-ribonucleic acid (miRNA), a primary micro-ribonucleic acid (pri-miRNA), a long non- coding RNA (lncRNA), a messenger ribonucleic acid (mRNA), a clustered regularly interspaced short palindromic repeats (CRISPR) related nucleic acid, a CRISPR-RNA (crRNA), a single guide ribonucleic acid (sgRNA), a trans-activating CRISPR ribonucleic acid (tracrRNA), a plasmid deoxyribonucleic acid (pDNA), a transfer ribonucleic acid (tRNA), an antisense oligonucleotide (ASO), an antisense ribonucleic acid (RNA), a guide ribonucleic acid, deoxy
  • siRNA
  • the first inlet may comprise a nucleic acid. In some embodiments, the first inlet may comprise mRNA. In some embodiments, the second inlet may comprise a nucleic acid. In some embodiments, the second inlet may comprise mRNA. In some embodiments, the third inlet may comprise a nucleic acid. In some embodiments, the third inlet may comprise mRNA. In some embodiments, the fourth inlet may comprise a nucleic acid. In some embodiments, the fourth inlet may comprise mRNA.
  • the mRNA is at least 15 mg (e.g., at least 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 80 mg, or 100 mg).
  • the lipid nanoparticle comprises one or more nucleic acids present in a weight ratio to the cationic ionizable lipid of from about 5:1 to about 1:100.
  • the weight ratio of the one or more nucleic acids to cationic ionizable lipid is at least 2:1, 1:1, 1:2, 1:5, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, or 1:50.
  • the weight ratio of the one or more nucleic acids to cationic ionizable lipids is at most 1:100, 1:90, 1:80, 1:70, 1:60, 1:50, 1:45, 1:40, 1:35, or 1:30. In some embodiments, the weight ratio of nucleic acid to cationic ionizable lipid is from about 5:1, 2.5:1, 1:1, 1:5, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:60, 1:70, 1:80, 1:90, or 1:100, or any range derivable therein. In some embodiments, the weight ratio is about 1:25 or about 1:7.
  • the weight ratio is from about 1:30 to about 1:40. In some embodiments, the weight ratio is about 1:30, 1:35, or about 1:40.
  • Inlet to Introduce Lipid Components any one of the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth inlet may be configured to introduce one or more lipid components of the lipid nanoparticle.
  • a lipid nanoparticle may comprise 2, 3, 4, 5, 6 or more components.
  • a lipid nanoparticle comprises 2 components.
  • a lipid nanoparticle comprises 3 components.
  • a lipid nanoparticle comprises 4 components.
  • a lipid nanoparticle comprises 5 components.
  • a lipid nanoparticle comprises 6 (e.g., or more) components.
  • a lipid nanoparticle comprises one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, and a PEGylated lipid.
  • a lipid nanoparticle comprises a nucleic acid and one or more lipid components (e.g., a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, and a PEGylated lipid).
  • the first inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • a therapeutic agent e.g., a nucleic acid
  • a cationic ionizable lipid e.g., a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • the third inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • the fourth inlet may be configured to introduce one or Atty Dkt No.: 58530-742601 more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • the fifth inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • a therapeutic agent e.g., a nucleic acid
  • a cationic ionizable lipid e.g., a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • the seventh inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • a therapeutic agent e.g., a nucleic acid
  • a cationic ionizable lipid e.g., a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • the ninth inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • the tenth inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid.
  • the lipid nanoparticles provided herein comprise a cationic ionizable lipid.
  • Cationic ionizable lipids as described elsewhere in the field may be used herein.
  • examples of cationic ionizable lipids may be found at U.S. Patent No.11,542,229, U.S. Patent No.11,247,968, U.S. Patent No.11,766,408, and U.S. Patent No.11,229,609, each of which are incorporated by reference herein in their entireties.
  • the cationic ionizable lipid is present in a molar percentage of the total lipid components of from about 2% to about 60%. In some embodiments, the molar percentage of the cationic ionizable lipid is from about 5% to about 50%.
  • the molar percentage of the cationic ionizable lipid is from about 5% to about 30%. In some embodiments, the molar percentage of the cationic ionizable lipid is from about 7.5% to about 20%. In some embodiments, the molar percentage of the cationic ionizable lipid is at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the molar percentage of the cationic ionizable lipid is at most 70%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, or 25%. [0090]
  • the lipid nanoparticles may comprise a steroid or steroid derivative.
  • the steroid or steroid derivative comprises any steroid or steroid derivative.
  • the term “steroid” is a class of compounds with a four ring Atty Dkt No.: 58530-742601 17 carbon cyclic structure which can further comprises one or more substitutions including alkyl groups, alkoxy groups, hydroxy groups, oxo groups, acyl groups, or a double bond between two or more carbon atoms. Examples of steroids may be found at U.S. Patent No.11,542,229, U.S. Patent No.11,247,968, U.S. Patent No.11,766,408, and U.S.
  • the steroid or steroid derivative is present in a molar percentage of the total lipid components of from about 20% to about 60%. In some embodiments, the steroid or steroid derivative is present in a molar percentage of from about 30% to 50%. In some embodiments, the steroid or steroid derivative is present in a molar percentage of from about 39% to about 46%. In some embodiments, the steroid or steroid derivative is present in a molar percentage of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, or 55%.
  • the steroid or steroid derivative is present in a molar percentage of at most 60%, 55%, 50%, 45%, 40%, 35%, 30%, or 25%.
  • the lipid nanoparticles comprise PEG or a PEGylated lipid.
  • the present disclosure comprises using any lipid to which a PEG group has been attached. Examples of PEG or a PEGylated lipids may be found at U.S. Patent No. 11,542,229, U.S. Patent No.11,247,968, U.S. Patent No.11,766,408, and U.S. Patent No. 11,229,609, each of which are incorporated by reference herein in their entireties.
  • the PEG or PEGylated lipid is present in a molar percentage of the total lipid components of from about 0.1% to about 20%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of from about 0.5% to about 10%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of from about 0.5% to about 5%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of from about 2% to about 2.8%.
  • the PEG or PEGylated lipid is present at a molar percentage of at least 0.1%, 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, or 8%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of at most 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, or 2%.
  • the lipid nanoparticles comprise a phospholipid.
  • the polymers are mixed with one or more phospholipids to create a composition. In some embodiments, any lipid which also comprises a phosphate group.
  • the phospholipid is a structure which contains one or two long chain C6-C24 alkyl or alkenyl groups, a glycerol or a sphingosine, one or two phosphate groups, and, optionally, a small organic molecule.
  • Examples of phospholipids may be found at U.S. Patent No.11,542,229, Atty Dkt No.: 58530-742601 U.S. Patent No.11,247,968, U.S. Patent No.11,766,408, and U.S. Patent No.11,229,609, each of which are incorporated by reference herein in their entireties.
  • the phospholipid is present in a molar percentage of total lipid components of about 1% to about 40%. In some embodiments, the phospholipid is present in a molar percentage of total lipid components of about 10% to about 40%. In some embodiments, the phospholipid is present in a molar percentage of total lipid components of at least about 5%, 7.5%, 10%, 15%, 17%, 18%, 20%, 22%, or 25%. In some embodiments, the phospholipid is present in a molar percentage of total lipid components of at most about 40%, 38%, 35%, 33%, 30%, 28%, 25%, 22%, or 20%.
  • the lipid nanoparticles comprise one or more selective organ targeting (SORT) compounds, which lead to the selective delivery of the lipid nanoparticle to a particular organ.
  • SORT selective organ targeting
  • This compound may be a lipid, a small molecule therapeutic agent, a sugar, a vitamin, or a protein.
  • SORT lipids can be found at U.S. Patent No.11,766,408 and U.S. Patent No.11,229,609, which are incorporated by reference herein in their entireties.
  • the one or more SORT compounds are present in a molar percentage of total lipid components of from about 5% to about 50%.
  • the one or more SORT compounds are present in molar percentage of from about 10% to about 45%. In some embodiments, the one or more SORT compounds are present in an amount of about 20% to about 40%. In some embodiments, the one or more SORT compounds are present in an amount of at least 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 50%. In some embodiments, the one or more SORT compounds are present in an amount of at most 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10%. [0098] In some aspects, the present disclosure provides one or more lipids with one or more hydrophobic components and a permanently cationic group.
  • the permanently cationic lipid may contain a group which has a positive charge regardless of the pH. Permanently cationic lipids as described elsewhere in the field may be used. In some instances, examples of permanently cationic groups can be found at U.S. Patent No.11,766,408 and U.S. Patent No.11,229,609, which are incorporated by reference herein in their entireties. [0099] In some embodiments, the lipid nanoparticle may comprise a permanently cationic lipid at any suitable amount. In some instances, examples of permanently cationic lipids can be found at U.S. Patent No.11,766,408 and U.S. Patent No.11,229,609, which are incorporated by reference herein in their entireties.
  • the present disclosure provides one or more lipids with one or more hydrophobic components and a permanently anionic group.
  • examples of permanently anionic lipids can be found at U.S. Patent No.11,766,408 and U.S. Patent No. 11,229,609, which are incorporated by reference herein in their entireties.
  • Static Mixer [00101] The present disclosure provides systems comprising a static mixer. In some embodiments, the static mixer may be downstream of the conduit. In some embodiments, the static mixer may be upstream of the conduit. In some embodiments, the static mixer may be parallel to the conduit. [00102] The system may comprise a static mixer.
  • the system may comprise a static mixer, wherein the nucleic acid and one or more lipid components are mixed to generate a mixture fluid, and wherein the nucleic acid and the one or more lipid components may not contact each other prior to entering the junction site. In some embodiments, the nucleic acid and one or more lipid components are mixed to generate a mixture fluid within the static mixer.
  • a static mixer described herein may comprise a design as illustrated in FIG. 29A, FIG.29B, or FIG.29C.
  • a static mixer described herein may comprise a design as illustrated in FIG.29A, FIG.29B, FIG.29C, or a combination thereof.
  • a static mixer described herein may comprise a design as illustrated in FIG.29A.
  • a static mixer described herein may comprise a design as illustrated in FIG.29B.
  • a static mixer described herein may comprise a design as illustrated in FIG.29C.
  • a static mixer described herein may comprise design A1, as illustrated in FIG. 29A.
  • a static mixer described herein may comprise design A2, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design A3, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design A4, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design A5, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design A6, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design B1, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design B2, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design B3, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design B4, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design C1, as illustrated in FIG. 29A.
  • a static mixer described herein may comprise design C2, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design C3, as illustrated in FIG.29A.
  • a static mixer described herein may comprise design C4, as illustrated in FIG.29B.
  • a static mixer described Atty Dkt No.: 58530-742601 herein may comprise design C5, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C6, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C7, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C8, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C9, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C10, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C11, as illustrated in FIG. 29B.
  • a static mixer described herein may comprise design C12, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C13, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C14, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C15, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C16, as illustrated in FIG.29B.
  • a static mixer described herein may comprise design C17, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design C18, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design C18, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D1, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D2, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D3, as illustrated in FIG. 29C.
  • a static mixer described herein may comprise design D4, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D5, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D6, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D7, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D8, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D9, as illustrated in FIG.29C.
  • a static mixer described herein may comprise design D10, as illustrated in FIG.29C.
  • a static mixer described herein may comprise a design of A1, A2, A3, A4, A5, A6, B1, B2, B3, B4, C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 C11, C12, C13, C14, C15, C16, C17, C18, D1, D2, D3, D4, D5, D6, D7, D8, D9, D10, or any combination thereof.
  • the static mixer may have a length less than about 1000 millimeter (mm), 500 mm, 300 mm, 200 mm, 100 mm, 90 mm, 80 mm, 70 mm, 60 mm, 50 mm, 45 mm, 40 mm, 35 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 9 mm, 8 mm, 7 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, or 1 mm.
  • the static mixer comprises a length of at least 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 15 mm, 20 mm, 25 mm, or 30 mm.
  • the static mixer comprises a length of from about 5 mm to about 80 mm. In some Atty Dkt No.: 58530-742601 embodiments, the static mixer comprises a length of from about 5 mm to about 40 mm. In some embodiments, the static mixer comprises a length of from about 10 mm to about 35 mm. In some embodiments, the static mixer comprises a length of about 20 mm, as shown in FIGs.18A-18C. In some embodiments, the static mixer comprises a length of about 30 mm, as shown in FIGs. 19A-19C. In some embodiments, the static mixer comprises a length of about 25 mm, as shown in FIGs.20A-20C.
  • the static mixer comprises a length of about 12 mm, as shown in FIGs.22A-C.
  • the static mixer may have a length less than about 10 inches (in), 5 in, 4 in, 3 in, 2 in, 1.5 in, 1.2 in, 1 in, 0.9 in, 0.8 in, 0.7 in, 0.6 in, 0.5 in, 0.4 in, 0.3 in, 0.2 in, or 0.1 in.
  • the static mixer has a length of about 4.8 mm.
  • the static mixer may have a length about 0.8 in, as shown in FIGs. 18A-18C.
  • the static mixer may have a length about 1.2 in, as shown in FIGs.19A-19C.
  • the static mixer may have a length of about 1 in. [00107] In some embodiments, the static mixer may have an inner diameter of less than about 5 in, 4 in, 3 in, 2 in, 1 in, 3 ⁇ 4 in, 1 ⁇ 2 in, 3/8 in, 1 ⁇ 4 in, 3/16 in, 1/8 in, 1/16 in, or 1/32 in. In some embodiments, the static mixer comprises an inner diameter of less than about 127 mm, 102 mm, 76 mm, 50 mm, 25 mm, 19 mm, 13 mm, 9.5 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, 1.5 mm, 1 mm, or 0.75 mm.
  • the static mixer comprises an inner diameter of at least 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 4 mm, or 5 mm. In some embodiments, the static mixer comprises an inner diameter of from about 0.5 mm to about 20 mm. In some embodiments, the static mixer comprises an inner diameter of from about 1 mm to about 10 mm. In some embodiments, the static mixer comprises an inner diameter of from about 1 mm to about 5 mm. In some embodiments, the static mixer comprises an inner diameter of about 3 mm.
  • the static mixer may have 1 element, 2 or more elements, 4 or more elements, 6 or more elements, 8 or more elements, 12 or more elements, 16 or more elements, 24 or more elements, or 36 or more elements.
  • the static mixer comprises at least 1 element, at least 2 elements, at least 3 elements, at least 5 elements, at least 8 elements, at least 12 elements, or at least 20 elements.
  • the static mixer comprises at most 54 elements, at most 48 elements, at most 36 elements, at most 24 elements, at most 16 elements, or at most 12 elements.
  • the static mixer comprises from 1 to 12 elements. In some embodiments, the static mixer comprises from 5 to 10 elements. In some embodiments, the static mixer comprises 8 elements.
  • the systems provided herein further comprise a splitter insert, which may be arranged within the system in various ways.
  • the static Atty Dkt No.: 58530-742601 mixer is inserted into the splitter insert, as shown in FIG.8A and FIG.8B.
  • the elements of the static mixer may be arranged in various ways.
  • a splitter insert may be inserted in the static mixer to split the flows prior to mixing the elements and to ensure consistent mixing, as shown in FIG.8A and 8B.
  • the splitter insert may have a length of less than about 70 mm, 50 mm, 40 mm, 35 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 5 mm, or 1 mm. In some embodiments, the splitter insert comprises a length of at least 1 mm, 5 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 35 mm, or 40 mm. In some embodiments, the splitter insert comprises a length of from about 1 mm to about 50 mm, from about 10 mm to about 40 mm, from about 20 mm to about 40 mm. In some embodiments, the splitter insert comprises a length of from about 30 mm to about 40 mm.
  • the splitter insert comprises a length of about 35 mm, as shown in FIG. 8A and FIG.8B.
  • the static mixer comprises a splitter insert of FIG.8A or FIG.8B.
  • the splitter insert is a tee-shaped conduit.
  • the splitter insert is a Y-shaped conduit.
  • the splitter insert is an L-shaped conduit.
  • any suitable number of elements in a static mixer may be inserted into a splitter insert, based on for example, the length of the elements and the length of the splitter insert. In some embodiments, at least 1 of the elements of the static mixer is inserted into the splitter insert.
  • At least 2, 3, 4, 5, 6, 7, or 8 of the elements of the static mixer may be inserted into the splitter insert. In some embodiments, at most 75% of the elements of the static mixer may be inserted into the splitter insert. In some embodiments, at most 50% of the elements of the static mixer may be inserted into the splitter insert. In some embodiments, at most 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 of the elements of the static mixer may be inserted into the splitter insert. [00113] In some embodiments, any suitable number of elements in a static mixer may be inserted into a tee-shaped conduit, based on for example, the length of the elements and the length of the tee-shaped conduit.
  • At least 1 of the elements of the static mixer is inserted into the tee-shaped conduit. In some embodiments, at least 2, 3, 4, 5, 6, 7, or 8 of the elements of the static mixer may be inserted into the tee-shaped conduit. In some embodiments, at most 75% of the elements of the static mixer may be inserted into the tee- shaped conduit. In some embodiments, at most 50% of the elements of the static mixer may be Atty Dkt No.: 58530-742601 inserted into the tee-shaped conduit. In some embodiments, at most 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 of the elements of the static mixer may be inserted into the tee-shaped conduit.
  • 2 of the 8 or more elements in a static mixer may be inserted into a tee-shaped conduit prior to the static mixer.
  • 6 of the 8 or more elements in a static mixer may be downstream from an outlet in a tee-shaped conduit.
  • 4 of the 12 or more elements in a static mixer may be inserted into a tee-shaped conduit prior to the static mixer.
  • 8 of the 12 or more elements in a static mixer may be downstream from an outlet in the tee-shaped conduit.
  • 2 of the 16 or more elements in a static mixer may be inserted into a tee-shaped conduit prior to the static mixer.
  • 4 of the 16 or more elements in a static mixer may be inserted into a tee-shaped conduit prior to the static mixer.
  • 12 of the 16 or more elements in a static mixer may be downstream from an outlet in the tee-shaped conduit.
  • 14 of the 16 or more elements in a static mixer may be downstream from an outlet in the tee-shaped conduit.
  • any suitable number of elements in a static mixer may be inserted into a Y-shaped conduit, based on for example, the length of the elements and the length of the Y-shaped conduit. In some embodiments, at least 1 of the elements of the static mixer is inserted into the Y-shaped conduit.
  • At least 2, 3, 4, 5, 6, 7, or 8 of the elements of the static mixer may be inserted into the Y-shaped conduit. In some embodiments, at most 75% of the elements of the static mixer may be inserted into the Y-shaped conduit. In some embodiments, at most 50% of the elements of the static mixer may be inserted into the Y-shaped conduit. In some embodiments, at most 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 of the elements of the static mixer may be inserted into the Y-shaped conduit. [00116] In some embodiments, 2 of the 8 or more elements in a static mixer may be inserted into a Y-shaped conduit prior to the static mixer.
  • 6 of the 8 or more elements in a static mixer may be downstream from an outlet in the Y-shaped conduit.
  • 4 of the 12 or more elements in a static mixer may be inserted into a Y-shaped conduit prior to the static mixer.
  • 8 of the 12 or more elements in a static mixer may be downstream from an outlet in the Y-shaped conduit.
  • 2 of the 16 or more elements in a static mixer may be inserted into a Y-shaped conduit prior to the static mixer.
  • 4 of the 16 or more elements in a static mixer may be inserted into a Y-shaped conduit prior to the static mixer.
  • 12 of the 16 or more elements in a static mixer may be downstream from an outlet in the Y-shaped conduit.
  • 14 of the 16 or more elements in a static mixer may be downstream from an outlet in the Y-shaped conduit.
  • any suitable number of elements in a static mixer may be inserted into a L-shaped conduit, based on for example, the length of the elements and the length of the L-shaped conduit.
  • at least 1 of the elements of the static mixer is inserted into the L-shaped conduit.
  • at least 2, 3, 4, 5, 6, 7, or 8 of the elements of the static mixer may be inserted into the L-shaped conduit.
  • At most 75% of the elements of the static mixer may be inserted into the L-shaped conduit. In some embodiments, at most 50% of the elements of the static mixer may be inserted into the L-shaped conduit. In some embodiments, at most 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 of the elements of the static mixer may be inserted into the L-shaped conduit. [00118] In some embodiments, 2 of the 8 or more elements in a static mixer may be inserted into a L-shaped conduit prior to the static mixer. In some embodiments, 6 of the 8 or more elements in a static mixer may be downstream from an outlet in the L-shaped conduit. In some embodiments, 4 of the 12 or more elements in a static mixer may be inserted into a L-shaped conduit prior to the static mixer.
  • 8 of the 12 or more elements in a static mixer may be downstream from an outlet in the L-shaped conduit.
  • 2 of the 16 or more elements in a static mixer may be inserted into a L-shaped conduit prior to the static mixer.
  • 4 of the 16 or more elements in a static mixer may be inserted into a L-shaped conduit prior to the static mixer.
  • 12 of the 16 or more elements in a static mixer may be downstream from an outlet in the L-shaped conduit.
  • 14 of the 16 or more elements in a static mixer may be downstream from an outlet in the L-shaped conduit.
  • each element of the static mixer may have a length less than about 2 in, 1.5 in, 1.2 in, 1 in, 0.9 in, 0.8 in, 0.7 in, 0.6 in, 0.5 in, 0.4 in, 0.3 in, 0.2 in, 0.19 in, 0.18 in, 0.17 in, 0.16 in, 0.15 in, 0.14 in, 0.13 in, 0.12 in, 0.11 in, or 0.10 in.
  • each element of the static mixer comprises a length of less than 50 mm, 40 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 5 mm, or 2.5 mm.
  • each element of the static mixer comprises a length of at least 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 5 mm, or 6 mm. In some embodiments, each element of the static mixer comprises a length of from about 1 mm to about 5 mm. In some embodiments, each element of the static mixer comprises a length of from about 1 mm to about 10 mm. In some embodiments, each element of the static mixer comprises a length of about 2.5 mm, as shown in FIGs.18A-C. In some Atty Dkt No.: 58530-742601 embodiments, each element of the static mixer comprises a length of about 3.8 mm, as shown in FIGs.19A-C.
  • each element of the static mixer comprises a length of about 3.2 mm, as shown in FIGs.21A-C. In some embodiments, each element of the static mixer comprises a length of about 1.5 mm, as shown in FIGs.22A-C. In some embodiments, each element of the static mixer comprises a length of about 4.8 mm, as shown in FIGs.23A-C. [00120] In some embodiments, each element of the static mixer may have a 45° twist. In some embodiments, each element of the static mixer may have a 90° twist, as shown in FIGs.22A- 22C. In some embodiments, each element of the static mixer may have a 135° twist.
  • each element of the static mixer may have a 180° twist, as shown in FIGs.9A-9C, 25 and 28. In some embodiments, each element of the static mixer may have a 225° twist. In some embodiments, each element of the static mixer may have a 270° twist, as shown in FIGs. 23A-23C, 26, and 27. In some embodiments, each element of the static mixer may have a 315° twist. In some embodiments, each element of the static mixer may have a -45° twist. In some embodiments, each element of the static mixer may have a -90° twist. In some embodiments, each element of the static mixer may have a -135° twist.
  • each element of the static mixer may have a -180° twist. In some embodiments, each element of the static mixer may have a -225° twist. In some embodiments, each element of the static mixer may have a - 270° twist. In some embodiments, each element of the static mixer may have a -315° twist. [00121] In some embodiments, each element of the static mixer makes a 270° twist every 1/8 inch. [00122] In some embodiments, the static mixer may have a 90° turn mixer, as shown in FIGs. 22A-22C. In some embodiments, the static mixer may have a 270° turn mixer, as shown in FIGs.23A-23C.
  • each element of the static mixer may have a cross section thickness less than about 5 mm, 4 mm, 3 mm, 2 mm, 1 mm, 0.5 mm, 0.4 mm, 0.3 mm, 0.2 mm, or 0.1 mm.
  • each element of the static mixer comprises a cross section thickness of at least 0.1 mm, 0.2 mm, 0.3 mm, 0.5 mm, 0.75 mm, 1 mm, 1.1 mm, 1.2 mm, 1.3 mm, 1.4 mm, or 1.5 mm.
  • each element of the static mixer comprises a cross section thickness of from about 0.1 mm to about 2 mm.
  • each element of the static mixer comprises a cross section thickness of from about 1 mm to about 2 mm. In some embodiments, each element of the static mixer comprises a cross section thickness of about 1 mm. In other embodiments, each element of the static mixer comprises a cross section thickness of about 1 mm. Atty Dkt No.: 58530-742601 [00124]
  • the static mixer may have one or more channel mixers. In some embodiments, the static mixer may have a 3-channel mixer. In some embodiments, the 3- channel mixer may be Y-shaped, as shown in FIGs.20A-20C. In some embodiments, the static mixer may have a 4-channel mixer.
  • the 4-channel mixer may be X- shaped, as shown in FIGs.21A-21C.
  • the systems herein provide for the requirement of reduced pressures as opposed to alternative systems, such as cross mixing systems. Reduced pressures may be useful to decrease the imposition of shear stress on the lipid nanoparticles and result in lower backpressure on pumping systems. Reduced pressures may be useful to allow adoption into larger scale production. In some embodiments, the pressure may be backpressure. In some embodiments, the systems provided herein reach a pressure of no more than 8 pounds per square inch gauge (psig), whereas cross mixing systems may reach a pressure of over 20 psig (e.g., 23.3 psig as described in the Examples).
  • psig pounds per square inch gauge
  • cross mixing systems may reach a pressure of over 20 psig (e.g., 23.3 psig as described in the Examples).
  • the pressure of a system provided reaches no more than 20 psig (e.g., no more than 18 psig, 17 psig, 16 psig, 15 psig, 14 psig, 13 psig, 12 psig, 11 psig, 10 psig, 9 psig, or 8 psig). In some embodiments, the pressure of the system reaches at least 3 psig (e.g., at least 4 psig, 5 psig, 8 psig, 10 psig, 12 psig, 14 psig, or 16 psig). In some embodiments, the pressure of the systems provided herein reach from about 3 psig to about 20 psig.
  • the pressure of the systems provided herein reach from about 5 psig to about 15 psig, about 5 psig to about 10 psig, or about 6 psig to about 9 psig.
  • Channel [00126]
  • the present disclosure provides systems comprising a channel housing the static mixer.
  • the channel may be downstream of the conduit.
  • the channel may be upstream of the conduit.
  • the channel may be parallel to the conduit.
  • the system may comprise a channel.
  • the system may comprise a channel, wherein the channel houses a static mixer.
  • the system may comprise a channel, wherein the channel houses an element of a static mixer.
  • the system may comprise a channel, wherein the channel houses a plurality of elements of a static mixer.
  • the system may comprise a channel, wherein the channel houses an element.
  • the system may comprise a channel, wherein the channel houses a plurality of elements, In some embodiments, the channel provides a counterflow.
  • the system may comprise a channel, wherein the nucleic acid and one or more lipid components are mixed to generate a mixture fluid. In some embodiments, the nucleic acid and one or more lipid components are mixed to generate a mixture fluid within the channel.
  • the channel may have a length less than about 1000 millimeter (mm), 500 mm, 300 mm, 200 mm, 100 mm, 90 mm, 80 mm, 70 mm, 60 mm, 50 mm, 45 mm, 40 mm, 35 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 9 mm, 8 mm, 7 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, or 1 mm.
  • mm millimeter
  • the channel comprises a length of at least 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 15 mm, 20 mm, 25 mm, or 30 mm. In some embodiments, the channel comprises a length of from about 5 mm to about 80 mm. In some embodiments, the channel comprises a length of from about 5 mm to about 40 mm. In some embodiments, the channel comprises a length of from about 10 mm to about 35 mm.
  • the channel may have a length less than about 10 inches (in), 5 in, 4 in, 3 in, 2 in, 1.5 in, 1.2 in, 1 in, 0.9 in, 0.8 in, 0.7 in, 0.6 in, 0.5 in, 0.4 in, 0.3 in, 0.2 in, or 0.1 in. In some embodiments, the channel has a length of about 4.8 mm. [00129] In some embodiments, the channel may have an inner diameter of less than about 5 in, 4 in, 3 in, 2 in, 1 in, 3 ⁇ 4 in, 1 ⁇ 2 in, 3/8 in, 1 ⁇ 4 in, 3/16 in, 1/8 in, 1/16 in, or 1/32 in.
  • the channel comprises an inner diameter of less than about 127 mm, 102 mm, 76 mm, 50 mm, 25 mm, 19 mm, 13 mm, 9.5 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, 1.5 mm, 1 mm, or 0.75 mm. In some embodiments, the channel comprises an inner diameter of at least 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 4 mm, or 5 mm. In some embodiments, the channel comprises an inner diameter of from about 0.5 mm to about 20 mm. In some embodiments, the channel comprises an inner diameter of from about 1 mm to about 10 mm.
  • the channel comprises an inner diameter of from about 1 mm to about 5 mm. In some embodiments, the channel comprises an inner diameter of about 3 mm. [00130] In some embodiments, the channel may be cylindrical, as shown in FIG. 25. In some embodiments, the channel may be square shaped, as shown in FIGs.26-28. In some embodiments, the channel may be rectangular shaped. In some embodiments, the channel may be any shaped.
  • the channel may have a twist at a length every less than about 2 in, 1.5 in, 1.2 in, 1 in, 0.9 in, 0.8 in, 0.7 in, 0.6 in, 0.5 in, 0.4 in, 0.3 in, 0.2 in, 0.19 in, 0.18 in, 0.17 in, 0.16 in, 0.15 in, 0.14 in, 0.13 in, 0.12 in, 0.11 in, or 0.10 in.
  • the channel may have a twist at a length every less than 50 mm, 40 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 5 mm, or 2.5 mm.
  • the channel may have a twist at a length every at least 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 5 mm, or 6 mm. In some embodiments, the channel may have a twist at a length every about 1 mm to about 5 mm. In Atty Dkt No.: 58530-742601 some embodiments, the channel may have a twist at a length every about 1 mm to about 10 mm. In some embodiments, the channel may have a twist at a length every 1/8 in. [00132] In some embodiments, the channel may have a plurality of twists. In some embodiments, the channel may have a 45° twist.
  • the channel may have a 90° twist. In some embodiments, the channel may have a 135° twist. In some embodiments, the channel may have a 180° twist. In some embodiments, the channel may have a 225° twist. In some embodiments, the channel may have a 270° twist. In some embodiments, the channel may have a 315° twist. In some embodiments, the channel may have a -45° twist. In some embodiments, the channel may have a -90° twist. In some embodiments, the channel may have a -135° twist. In some embodiments, the channel may have a -180° twist, as shown in FIG.26D, 27D, and 28D.
  • the channel may have a -225° twist. In some embodiments, the channel may have a -270° twist. In some embodiments, the channel may have a -315° twist. [00133] In some embodiments, the channel is square shaped and makes a -180° twist every 1/8 inch.
  • Pump [00134] The present disclosure provides systems comprising a pump.
  • the pump may be configured to provide one or more components of the lipid nanoparticle at a specified flow rate, such as a flow rate described elsewhere herein. In some embodiments, the pump is configured to provide the nucleic acid at a specified flow rate. In some embodiments, the pump is configured to provide the one or more lipids at a specified flow rate.
  • the pump is configured to provide the cationic ionizable lipid at a specified flow rate. In some embodiments, the pump is configured to provide the phospholipid at a specified flow rate. In some embodiments, the pump is configured to provide the polymer conjugated lipid at a specified flow rate. In some embodiments, the pump may be a peristaltic pump. In some embodiments, the pump may be a high performance liquid chromatography (HPLC) pump. In some embodiments, the pump may be a Levitronix pump. In some embodiments, the pump may be a Knauer pump. In some embodiments, the pump may be a Knauer BlueShadow pump.
  • HPLC high performance liquid chromatography
  • the present disclosure provides systems and methods for generating lipid nanoparticles.
  • Atty Dkt No.: 58530-742601 [00136]
  • the methods provided herein comprise providing a system, such as a system provided elsewhere herein.
  • the system comprises a conduit.
  • the conduit comprises a first inlet.
  • the conduit comprises a second inlet.
  • the conduit comprises a third inlet.
  • the conduit comprises a fourth inlet.
  • the conduit comprises an outlet.
  • the system comprises a static mixer.
  • the conduit comprises a channel.
  • the methods provided herein comprise introducing a nucleic acid to the first inlet.
  • the methods provided herein comprise introducing one or more lipid components of the lipid nanoparticle to the second inlet.
  • the systems or methods may comprise flowing a (e.g., first) solution comprising nucleic acid into a first inlet of a conduit, and a (e.g., second) solution comprising one or more lipid components into a second inlet of the conduit.
  • the first solution comprises any nucleic acid described elsewhere herein.
  • the second solution comprises one or more lipid components as described elsewhere herein.
  • the methods provided herein comprise introducing a nucleic acid to the first inlet or the second inlet or the third inlet or the fourth inlet. In some embodiments, the methods provided herein comprise introducing one or more lipid components of the lipid nanoparticle to the first inlet or the second inlet or the third inlet or the fourth inlet.
  • the systems or methods may comprise flowing a (e.g., first) solution comprising nucleic acid into a first inlet or a second inlet or a third inlet or a fourth inlet of a conduit, and a (e.g., second) solution comprising one or more lipid components into the first inlet or the second inlet or the third inlet or the fourth inlet of the conduit.
  • the first solution comprises any nucleic acid described elsewhere herein.
  • the second solution comprises one or more lipid components as described elsewhere herein.
  • the methods provided herein comprise joining the nucleic acid and the one or more lipid components at the outlet.
  • the first inlet and the second inlet may join into an outlet, such as an outlet described elsewhere herein, wherein the nucleic acid and the one or more lipid components are joined in the outlet.
  • the systems or methods may further comprise a static mixer.
  • the nucleic acid and the one or more lipid components are mixed within the static mixer to generate lipid nanoparticles.
  • the systems or methods may further comprise a static mixer, such as a static mixer described elsewhere herein, wherein Atty Dkt No.: 58530-742601 the nucleic acid and the one or more lipid components are mixed in a mixture fluid to generate lipid nanoparticles.
  • the systems or methods may further comprise a channel.
  • the nucleic acid and the one or more lipid components are mixed within the channel to generate lipid nanoparticles.
  • the systems or methods may further comprise a channel, such as a channel described elsewhere herein, wherein the nucleic acid and the one or more lipid components are mixed in a mixture fluid to generate lipid nanoparticles
  • the systems and methods may generate lipid nanoparticles with a low polydispersity index, wherein the polydispersity index may be no greater than about 0.2.
  • the systems and methods may generate lipid nanoparticles no greater than about 90 nanometers (nm).
  • the systems and methods may generate mixture fluids with a Reynolds number no greater than about 1000.
  • the systems and methods may have a ratio between a flow rate of the lipid component in the first inlet and a flow rate of the nucleic acid in the second inlet about 1:2.
  • the systems and methods may have an encapsulation efficiency no less than about 90%.
  • the conduit may comprise a first inlet, a second inlet, an outlet.
  • the conduit may further comprise a junction site downstream of the first inlet and the second inlet.
  • the method may comprise providing a static mixer.
  • the method may comprise providing a static mixer, wherein the static mixer is disposed at a distal end of the junction site.
  • the method may further comprise introducing a nucleic acid to the first inlet.
  • the method may further comprise introducing one or more lipid components of the lipid nanoparticle to the second inlet.
  • the method may further comprise joining the nucleic acid and the one or more lipid components at the outlet.
  • the method may further comprise using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid.
  • the method may further comprise using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid. Wherein the nucleic acid and the one or more lipid components may not contact each other prior to entering the junction site.
  • the conduit may comprise a first inlet, a second inlet, a third inlet, a fourth inlet, an outlet.
  • the method may comprise providing a static mixer.
  • the method may further comprise introducing a nucleic acid to the first inlet or the second inlet or the third inlet or the fourth inlet.
  • the method may further comprise introducing one or more lipid components of the lipid nanoparticle to the first inlet or the second inlet or the third inlet or the fourth inlet.
  • the method may further comprise joining the nucleic acid and the one or more lipid components at the outlet.
  • Atty Dkt No.: 58530-742601 The method may comprise providing a static mixer, wherein the static mixer may be configured to mix the nucleic acid and the one or more lipid components, wherein the static mixer comprises a plurality of elements, wherein each element of the static mixer makes a 270° twist every 1/8 inch.
  • the method may further comprise providing a channel housing the static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the channel is square shaped and makes a -180° twist every 1/8 inch.
  • the mixture fluid may have a polydispersity index (PDI) no greater than about 0.5, 0.4, 0.3, 0.25, 0.20, 0.19, 0.18, 0.17, 0.160.15, 0.14, 0.13, 0.12, 0.11, 0.10, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or 0,01.
  • PDI polydispersity index
  • the mixture fluid comprises a polydispersity index (PDI) of at least 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09.0.10, 0.11, 0.12, 0.13, 0.14, or 0.15.
  • the PDI may be about 0.05 to about 0.5.
  • the PDI may be about 0.05 to about 0.25.
  • the PDI may be about 0.05 to about 0.2.
  • the PDI may be about 0.1 to about 0.5.
  • the PDI may be about 0.1 to about 0.25.
  • the PDI may be about 0.1 to about 0.2.
  • the PDI may be about 0.05 to about 0.16. In some embodiments, the PDI may be about 0.09 to about 0.12.
  • the PDI may be measured using light scattering techniques, such as dynamic light scattering (DLS), or by electron microscopy (e.g., scanning electron microscopy or transmission electron microscopy).
  • the mixture fluid may have a Reynolds number of less than about 2500, 1500, 1200, 1000, 900, 800, 700, 600, 500, 400, 300, 200, or 100. In some embodiments, the mixture fluid may have a Reynolds number of at least 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000. In some embodiments, the Reynolds number is from about 100 to about 1200.
  • increasing ⁇ decreases the size of the Atty Dkt No.: 58530-742601 resulting lipid nanoparticles.
  • decreasing ⁇ the density of the fluid, increases the size of the resulting lipid nanoparticles.
  • an increase in a pressure difference increases ⁇ , the density of the fluid.
  • an increase in a pressure difference increases the Reynolds number.
  • a decrease in a pressure difference decreases ⁇ , the density of the fluid.
  • an increase in a pressure difference decreases the size of the resulting lipid nanoparticles.
  • a decrease in a pressure difference increases the size of the resulting lipid nanoparticles.
  • increasing u, the flow speed increases the Reynolds number. In some embodiments, decreasing u, the flow speed, decreases the Reynolds number. In some embodiments, modifying u, the flow speed, controls the size of the resulting lipid nanoparticle. In some embodiments, increasing u, the flow speed, decreases the size of the resulting lipid nanoparticles. In some embodiments, decreasing u, the flow speed, increases the size of the resulting lipid nanoparticles. In some embodiments, an increase in a pressure difference increases u, the flow speed. In some embodiments, an increase in a pressure difference increases the Reynolds number. In some embodiments, a decrease in a pressure difference decreases u, the flow speed.
  • a decrease in a pressure difference decreases the Reynolds number.
  • an increase in a pressure difference decreases the size of the resulting lipid nanoparticles.
  • a decrease in a pressure difference increases the size of the resulting lipid nanoparticles.
  • increasing L, the characteristic linear dimension increases the Reynolds number.
  • decreasing L, the characteristic linear dimension decreases the Reynolds number.
  • increasing L, the characteristic linear dimension decreases the size of the resulting lipid nanoparticles.
  • decreasing L, the characteristic linear dimension increases the size of the resulting lipid nanoparticles.
  • changing L, the characteristic linear dimension, and changing u, the flow speed maintain the Reynolds number.
  • L, the characteristic linear dimension is a channel size.
  • increasing ⁇ , the dynamic viscosity of the fluid decreases the Reynolds number.
  • decreasing ⁇ , the dynamic viscosity of the fluid increases the Reynolds number.
  • increasing ⁇ , the dynamic viscosity of the fluid increases the size of the resulting lipid nanoparticle.
  • decreasing ⁇ , the dynamic viscosity of the fluid decreases the size of the resulting lipid nanoparticle.
  • Atty Dkt No.: 58530-742601 [00152] In some embodiments, increasing the Reynolds number decreases the particle size of the lipid nanoparticle. In some embodiments, decreasing the Reynolds number increases the particle size of the lipid nanoparticle. In some embodiments, increasing the Reynolds number increases the particle size of the lipid nanoparticle. In some embodiments, decreasing the Reynolds number decreases the particle size of the lipid nanoparticle. In some embodiments, the Reynolds number remains the same as the particle size of the lipid nanoparticle increases. In some embodiments, the Reynolds number remains the same as the particle size of the lipid nanoparticle decreases.
  • the particle size (e.g., diameter) of the lipid nanoparticles decreases, such as shown in FIG.3A.
  • the PDI of the lipid nanoparticles remains about the same, such as shown in FIG.3B.
  • provided herein are methods of generating lipid nanoparticles, wherein the particle size is selected by increasing or decreasing the Reynolds number.
  • the method comprises increasing the Reynolds number to decrease particle size.
  • the method comprises decreasing the Reynolds number to increase particle size.
  • the relationship between the Reynolds number and particle size is an inverse relationship.
  • the lipid nanoparticle may have a size no greater than 500 nanometers (nm), 400 nm, 300 nm, 200 nm, 100 nm, 90 nm, 80 nm, 70 nm, 60 nm, 50 nm, 40 nm, 30 nm, 20 nm, 10 nm, 5 nm, or 1 nm.
  • the lipid nanoparticle comprise a size of at least 10 nm, 20 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 85 nm, 90 nm, 100 nm, 105 nm, 110 nm, 120 nm, 140 nm, 160 nm, 180 nm, or 200 nm.
  • the lipid nanoparticle comprises a size of from about 50 nm to about 300 nm.
  • the lipid nanoparticle comprises a size of from about 50 nm to about 100 nm.
  • the lipid nanoparticle comprises a size of from about 100 nm to about 200 nm.
  • the lipid nanoparticle size is measured by light scattering techniques (e.g., dynamic light scattering), or by electron microscopy (e.g., scanning electron microscopy or transmission electron microscopy).
  • particle size e.g., diameter
  • dilution comprises dilution with an aqueous solvent.
  • dilution comprises dilution with ethanol (e.g., such as to 12.5 v/v%).
  • the methods provided herein comprise introducing ethanol after the (e.g., first and second) solutions enter and leave the static mixer (e.g., post-mixer).
  • the amount particle size and polydispersity index (PDI) are influenced by the amount of ethanol introduced post mixer.
  • the amount of ethanol introduced post-mixer is at least 10%.
  • the amount of ethanol introduced post-mixer is at least about 12%, 15%, 17%, 25%, 33%, 40%, 45%, 50%, or at least 55%.
  • the characteristics of the resulting particles with varying amounts of ethanol introduced post-mixer are shown in Table 4.
  • the polydispersity is ⁇ 0.15 for lipid nanoparticles diluted with at least 25% ethanol. In some embodiments, the polydispersity is about 0.2 or higher for lipid nanoparticles diluted with l7% or less ethanol post- mixer. In some embodiments, the relationship between particle size and amount of ethanol is parabolic, with the lowest particle sizes exhibited after introduction of 25% ethanol post-mixer, such as described in Table 4. [00157] In some embodiments, an encapsulation efficiency may be no less than about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.
  • an encapsulation efficiency may be no more than about 99.9%, 99.5%, 99%, 98%, 96%, 94%, 90%, or 85%. In some embodiments, the encapsulation efficiency may be from about 40% to about 99.9%, about 50% to about 99%, about 60% to about 99%, about 70% to about 99%, about 80% to about 99%. In some embodiments the encapsulation efficiency is from about 90% to about 99%, such as shown in FIG.7C. [00158] In some embodiments, the lipid nanoparticles generated by the methods provided herein using the systems provided herein are stable for at least 24 hours (e.g., at least 48 hours, 1 week, 1 month, 3 months, or at least 6 months).
  • the lipid nanoparticles are stable for from about 24 hours to about 6 months, from about 24 hours to about 3 months, about 1 week to about 6 months, or from about 1 month to about 6 months. In some embodiments, the lipid nanoparticles are stable at a temperature of from about 2°C to about 8°C. In some embodiments, the lipid nanoparticles are stable at a temperature of about -80°C. [00159] In some embodiments, a time of mixing may be no less than about 5 milliseconds (ms), 10 ms, 15 ms, 20 ms, 25 ms, 30 ms, 35 ms, 40 ms, 45 ms, 50 ms, 55 ms, or 60 ms.
  • ms milliseconds
  • a mixing time may be no more than about 5 ms, 10 ms, 15 ms, 20 ms, 25 ms, 30 ms, 35 ms, 40 ms, 45 ms, 50 ms, 55 ms, or 60 ms. In some embodiments, the mixing time may be from about 5 ms to about 40 ms, such as shown in FIGs.17A-B. Atty Dkt No.: 58530-742601 [00160] While various embodiments of the disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the disclosure.
  • Example 1 Static Mixer for Lipid Nanoparticle Production
  • Cross-tee mixing is a mixing method that requires high energy input under turbulent flow conditions. Furthermore, when operating under high flow conditions, high back-pressure is exhibited, necessitating the use of specific pumps to manage the increased pressure. Alternative methods utilizing lower flow and back-pressure may be desirable to gain better control over the physicochemical properties of the drug product. Transitioning to lower flow and lower back- pressure regime could provide a broader selection of pump options and a better control on lipid nanoparticle (LNP) formation.
  • LNP lipid nanoparticle
  • Static mixers may prove to be a suitable choice for mixing processes.
  • Methods Lipid components for the desired formulation were dissolved in ethanol to achieve the appropriate concentration. For further details on lipid formulations, see Tables 8 and 11.
  • the aqueous mRNA phase consisted of 15 mM citrate pH 4 buffer with no mRNA added. For tests using mRNA, they were formulated according to Table 9.
  • Static mixers of varying length, diameter, and architecture were purchased from Cole Palmer, and are depicted in FIGs.1A-I. Each system was run with a 2:1, 3:1, or 5:1 buffer:lipid flow rate ratio (FRR) at varying total flow rate (TFR) corresponding to Reynolds numbers at static mixer outlet.
  • FRR lipid flow rate ratio
  • FIGs. 1A-I show schematics of each mixing system initially tested, with blue representing the organic lipid inlet and yellow representing the aqueous mRNA inlet.
  • Systems A and B both use the same mixing architecture, a Koflo 24-element 1/8” ID disposable static mixer with a polypropylene tube.
  • System C uses a Koflo 12-element 1 ⁇ 4” OD mixing element fitted into 1 ⁇ 4” ID platinum cured silicone tubing.
  • Systems D and E both are manipulated versions of system B, with system D (FIG.1D) having the mixing architecture pushed partway into the Y, and with system E (FIG.1E) having a makeshift flow splitter incorporated by way of a ziptie inserted into the Y.
  • FIG. 1G shows images of systems A, B, and C while being tested with food dye feedstocks.
  • the blue food dye is dissolved in ethanol, while the yellow food dye is dissolved in MilliQ water, to best illustrate lipid-mRNA mixing.
  • Images for systems A and B are shown at joint outlet Reynolds numbers of 25, 50, and 100, while system C is just shown at Re of 100.
  • the mixing quality of system C may be seen to be significantly lower than in systems A and B, which both seem to achieve a consistent green color after a few mixing elements at all tested Re. It is worth noting that clearance of air from the mixing systems was difficult, with bubbles Atty Dkt No.: 58530-742601 consistently travelling through the mixer through the entire process. This was more of an issue with the smaller-diameter mixer.
  • Table 2A and Table 2B shows all results from this day of testing, including two control runs conducted on the tee mixer (one at normal Re, another at Re comparable to most of the static mixer tests).
  • System B was tested again with higher citrate inline ratios (1:5 instead of 1:2), which resulted in larger PS than the previous tests at 1:2 FRR.
  • System B with 1:5 FRR and Re of 621 was also tested with immediate water dilution rather than 30-minute delayed water dilution, a test that showed PS and PDI independence to this change.
  • New systems with iterated designs were also tested, including systems F, G, and H, all at 1:2 FRR and Re of 621. Between these three systems, F and H both performed similarly well and better than any previously performed static mixer test.
  • FIGs.5A-B shows bar graphs of both the particle diameters post-dilution (FIG.5A) and PDI post-dilution (FIG.5B).
  • Day 4 System Runs [00178] Following the tests on Day 3, a test of higher ethanol amounts at static mixer outlet (pre-inline dilution) was conducted in order to find the optimum post-mixer ethanol concentration (ethanol (v/v) percentage) for the static mixing architecture.
  • System J (FIG.4A) was used for these tests, along with LNP 5 lipid and buffer, and a Reynolds number of approximately 1000 at mixing outlet was maintained across all tested conditions, assuming negligible change to Reynolds number by bulk viscosity and density (actual Re ranges from 920 to 1001).
  • PDI was generally low ( ⁇ 0.15) for formulations with at least 25% ethanol post-mixer, with 17% ethanol post-mixer resulting in PDI around 0.2.
  • Particle size showed a parabolic relationship to ethanol amount post-mixer, with sizes ranging from ca.85 to 160 nm and a minimum particle size reached around 33% ethanol post-mixer.
  • the 1:2 FRR being favorable aligns the static mixer with previous mixers, as this 1:2 ratio has previously been found to be a favorable mixing ratio for both tee and cross-tee mixers.
  • FIG.6A shows the particle diameters plotted against ethanol post-mixer
  • FIG.6B shows the PDI.
  • Table 5 [00181] Each formulation was made and processed through small-scale buffer exchange (PD 10 columns), then subsequent centrifuge-concentration (Amicon tubes), and filtration. Finally, storage stability was tested by storing aliquots at either 2-8°C or -80°C for over 24 hours. After each processing and storage step, particle size and size distribution were analyzed, the values being reported in Table 6A and Table 6B. Table 6A Table 6B Atty Dkt No.: 58530-742601 [00182] Particle sizes were consistently the lowest in the control system, with PS ranging from 61 to 65 nm, and PDI under 0.1.
  • the static mixer test with a FRR of 1:2 generally performed better than the 1:3 FRR, with the former achieving PDI consistently at or below 0.1 and PDI ranging from 72 to 77 nm.
  • the static mixer with 1:3 FRR interestingly had a particle size that steadily increased during processing, starting at 78 nm post-mix and increasing to 101 nm post-concentration. Post-concentration, the particle size increased by 14 nm.
  • Table 6A and Table 6B is the encapsulation efficiency, which was between 98 and 99% after both storage conditions for the control and 1:2 FRR static mixer conditions, and was much lower at 66-68% for the 1:3 FRR condition.
  • a second mixing system was constructed identical to System N (same tubing lengths, attachments, and connectors) but without any included static mixing elements. This was called the “No Mixer” system, and was tested to verify that the mixing was occurring primarily due to the static mixing elements and not due to the junction of fluids (e.g., via jet impingement).
  • the two systems were tested according to the flow parameters outlined in Table 7.
  • the samples were processed in the same manner as the samples previously prepared (Day 5 of testing), and the results from processing are shown in Table 8A and Table 8B.
  • Table 8A or Table 8B Not shown in Table 8A or Table 8B is the encapsulation efficiency post-storage for each formulation.
  • an encapsulation efficiency of 49-51% was achieved, whereas with the 1:2 FRR static mixing system an encapsulation of 96% was achieved. It was noted that the 96% encapsulation efficiency was lower than usually seen with this static mixing system, and lower than is usually expected with an LNP 5-CFTR formulation.
  • Table 8A Table 8B [00185] A general trend of lower performance can be seen in Table 8A and Table 8B for the control, no-mixer configuration. Particle sizes ranging from 186 to 235 nm were found during processing and post-storage, with maximum particle size seen after spin concentration and minimum particle size seen after refrigeration.
  • PDI for the no-mixer system ranged between 0.09 and 0.22, values generally being elevated compared to most mixing systems with LNP 5-CFTR.
  • the static mixer performed better in comparison to the no mixer system, with particle sizes ranging from 86 to 99 nm, with highest particle size seen post-mix and decreasing incrementally during processing.
  • the static mixer sample showed the smallest particle size post-refrigeration.
  • PDI was generally low for the static mixer sample, with values ranging from 0.06 to 0.14 and Atty Dkt No.: 58530-742601 peaking post-spin concentration. Once again, these values are not as ideal as those seen with the same formulation and conditions tested on previously (higher particle size and PDI, and lower encapsulation).
  • LNP 5-CFTR also resulted in large particles, but were smaller and less polydisperse than those of LNP 1-CFTR.
  • Particle sizes for LNP 5 ranged from 82 to 93 nm, with PDI ranging from 0.08 to 0.17 (the latter an outlier post-spin concentration). Smallest particle sizes were seen post-storage.
  • LNP 3-CFTR showed similar behavior in-process to LNP 5-CFTR, with particle size ranging from 82 to 95 nm, and PDI ranging from 0.08 to 0.13.
  • Both DNAI1 formulations performed better than all three CFTR formulations, with LNP 2- DNAI1 having particle size range of 78-86 nm and PDI range of 0.02-0.15.
  • LNP 4- DNAI1 had the lowest particle size range of the five formulations tested with a range of 63 to 73 nm and a PDI range of 0.06 to 0.16.
  • the static mixer system can indeed make viable, though slightly larger, nanoparticles with low polydispersity (less than 0.2) and moderate particle size (62-87 nm post-storage).
  • encapsulation efficiency values are not included in Table 10A and Table 10B, which were generally above 97%, with a few exceptions.
  • LNP 3-CFTR had a post-freeze/thaw encapsulation efficiency of 93%, which was peculiar since encapsulation efficiency post-refrigeration was 98.6%.
  • LNP 5-CFTR had an encapsulation efficiency of 94% post-refrigeration, which again was strange given its 97.8% post-freeze/thaw encapsulation efficiency.
  • LNP 2-DNAI1 had an encapsulation efficiency of ⁇ 90% after both storage methods.
  • Atty Dkt No.: 58530-742601 Table 10A Table 10B [00189]
  • the compared LNP 4 formulation uses a different mRNA (H1N1 rather than DNAI1), and LNP 1 uses a tagged version of CFTR.
  • the plotted comparisons are shown in FIGs.7A-C.
  • particle diameters are generally higher with the static mixing architecture than when made with the cross-tee architecture.
  • PDI is generally comparable between the two mixing systems, with static mixer having lower PDI in 5 of 9 of the comparisons.
  • Encapsulation Atty Dkt No.: 58530-742601 efficiency is also comparable, with the largest deviations in LNP 1 and LNP 2, with the former having higher encapsulation efficiency when made with static mixer, and the latter having lower encapsulation efficiency when made with static mixer.
  • Example 2 Static Mixer Mixing Simulation
  • a mixing simulation using the Reynolds stress model between ethanol and water demonstrated lower maximum shear stress and system pressure in the static mixer than in the cross mixer, as shown in FIGs.15A-C.
  • the static mixer had a length of 1 inch, and an inner diameter of about 1/8 inches.
  • the static mixer contained 8 elements, in which each element was about 1/8 inches in length, and each element makes an about 180° twist, as shown in FIG.15A.
  • FIG. 15A The cross-section thickness of each element was about 0.0394 inches (1 mm), as shown in FIG. 15A.
  • the ethanol flow rate was about 75.4 mL/min and the water flow rate was about 150.8 mL/min, as shown in FIG.15B.
  • FIGs.16A-B Shows the comparative system, a standard about 0.5 mm inner-diameter cross mixer fluid path with three inlets, two of which have water flow rates of about 40 mL/min and one of which has an ethanol flow rate of about 40 mL/min.
  • the outlet has an inner diameter of about 1/16 in. Both systems were simulated in Ansys Fluent, and the results were used to extract shear stress, total system pressure, and mass fraction of ethanol.
  • Cross sections were taken about every 0.1 mm perpendicular to direction of fluid travel, and on Atty Dkt No.: 58530-742601 these cross-sections area-weighted average shear stress and standard deviation of ethanol mass fraction were calculated.
  • the average shear stress term was used to evaluate the amount of shear stress each mixing architecture imposed on the mixing fluids, and the standard deviation of ethanol mass fraction was used to evaluate the homogeneity of the fluid, with low standard deviation representing high degree of mixing/homogeneity.
  • Time was calculated in milliseconds from the fluid flow rates from each system and the cross-sectional areas of each evaluated cross- section. A time point of zero was set at the initial point of water-ethanol flow intersection for each mixing system.
  • FIGs.17A-C The results from these simulations are shown in FIGs.17A-C. Comparing ethanol mass fraction standard deviation results from the two different mixers (FIG.17A), the cross mixer is shown to have a faster decrease in standard deviation, indicating a faster mixing process when compared to the simulated static mixing system. However, when comparing area-averaged shear stress (FIG.17B), the cross mixer is shown to have much higher shear stress during the mixing process, with a maximum reaching almost about 1000 Pa, which is over five times the maximum shear stress reached by the static mixing system.
  • Loctite 3955, FST, Black was selected as the mixer material, known for its chemical resistance to ethanol. FST composite resins are fire, smoke, and toxicity resistant.
  • a Koflo analogue design (Design 1, No. A1) was made, modeled after the Koflo static mixers.
  • the interior elements are helical elements with 180° twists (FIG.25A).
  • the exterior housing comprises 2 inlets with 1/8 inch barbs that are 120° apart (FIG.25B), which is wider than the typical Y connection of a 60° separation to allow space for tubing attachments, and zip ties.
  • the channel is cylindrical shape which houses the elements (FIG.25C).
  • Atty Dkt No.: 58530-742601 Another design (Design 2, No. D10) was made, which included helical elements with 270° twists (FIG.26A).
  • the exterior housing comprises 2 inlets (FIG.26B) and the channel is square shaped (FIG.26C) and comprise -180° twists (FIG.26D).
  • Another design (Design 3, No. D7) was made, which included helical elements with 270° twists (FIG.27A).
  • the exterior housing comprises 4 inlets (FIG.27B) and the channel is square shaped (FIG.27C) and comprise -180° twists (FIG.27D).
  • Another design Design 5, No.
  • a cutoff back-pressure of 10 psig was used to narrow down results, as this is 2/3 of the maximum rated pressure of a low- shear pump system we hope to use to drive the mixer flow. This cutoff was used to account for pressure increases from extra tubing as well as possible unquantified inaccuracy of the CFD results.
  • Design No. D7 had the highest simulated mixing speed as well as a low max average shear stress, making it a lead candidate.
  • Design No. D6 was also considered due to its similar design to Design No. D7, but with almost 2 psi lower simulated back-pressure.
  • Design No. D10 was considered due to its identical mixer design to Design No. D7 but with two inlets rather than four, thereby simplifying upstream connections.
  • Design No. A1 were successfully manufactured and tested.
  • Design No. D7 performed best at lower flow rates and was chosen to be tested at the higher simulated total flow rate of ⁇ 905 mL/min.
  • the back-pressure was measured to be approximately 15 psi, which is significantly higher than the simulated 6 psi back-pressure, but this was expected to a degree since the simulated back-pressure did not include contributions from upstream or downstream tubing and connections.
  • Example 4 Manufactured Static Mixer Initial Tests [00204] Each of the selected manufactured static mixers were tested against a cross mixer and a tee mixer, all at a 3:1 buffer to lipid ratio (empties).
  • a higher buffer pH can decrease particle size, so citrate buffer from pH 4 to pH 6 were tested. Buffer strength may also have an effect on particle size, so citrate buffer strength will be varied as well as adding sodium chloride (NaCl) to change charge screening and encourage particle formation. All conditions and tests performed are shown in Table 14 for empty LNPs with a FRR (Lipid:mRNA) ratio of 1:3. Table 14 Atty Dkt No.: 58530-742601 *Reynolds numbers based on initial SM control rates [00205] The initial tests (Tests 1-7) and control comparison results are shown in Table 15.
  • Knauer BlueShadow 80P LC pumps 1000 mL/min was used to achieve the required flow rates, and different parameters as were tested on peristaltic pumps.
  • the simulated flow parameters were 904.8 total flow rate (mL/min), max back-pressure of 5.77 psig, and a mixing efficiency of 224% as compared to the original static design.
  • Table 19 shows the test conditions on the HPLC pumps, which were set to mimic the test conditions on the peristaltic pump.
  • Table 19 [00214] Tests were performed with RTX0051-Empty formulation at different total flow rates. As shown in Table 20, the four tested conditions resulted in smaller particles than those made with the peristaltic setup.
  • the particle size ranged from 60-75 nm post-PD-10, with the highest flow condition resulting in the best particles across all sampling regimes. Minimal trend could be seen with regards to total flow rate, with all conditions creating decent empty particles.
  • Table 20 Atty Dkt No.: 58530-742601 [00215] As the results indicate, the HPLC pumps generated particles were much smaller. Peristaltic fluctuations may have a major effect on mixing efficiency at high flow rates. With the Design No. D7 mixer, sub-65 nm empty particles were achieved as shown in Table 20. This demonstrated a 20 nm reduction in size compared to the Koflo analogue, Design No. A1, which generated ⁇ 84 to 87 nm particles as shown in Table 15.
  • Embodiment 1 A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii.
  • a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; Atty Dkt No.: 58530-742601 i ii. an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and ( b) a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is no greater than about 0.25.
  • Embodiment 2. The system of embodiment 1, wherein the conduit is tee-shaped.
  • Embodiment 3. The system of embodiment 1, wherein the conduit is Y-shaped.
  • Embodiment 5 The system of embodiment 1, wherein the conduit is L-shaped.
  • Embodiment 6. The system of any one of the preceding embodiments, wherein the static mixer is less than about 40 millimeters (mm) in length.
  • Embodiment 7. The system of any one of the preceding embodiments, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 8. The system of any one of the preceding embodiments, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 9 The system of any one of the preceding embodiments, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 10. The system of any one of the preceding embodiments, wherein the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch.
  • Embodiment 11 The system of embodiment 10, wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 12. The system of embodiment 10, wherein the inner diameter is about 1/8 inch.
  • Embodiment 13 The system of any one of the preceding embodiments, wherein the polydispersity index is about 0.05 to about 0.25.
  • Embodiment 14 The system of embodiment 13, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 15 The system of any one of embodiments 1-12, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 16 The system of any one of the preceding embodiments, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Atty Dkt No.: 58530-742601 Embodiment 17.
  • the system of embodiment 16, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 18. The system of embodiment 16, wherein the Reynolds number is about 1000.
  • Embodiment 19. The system of any one of the preceding embodiments, wherein the lipid nanoparticle is no greater than about 100 nanometers (nm).
  • Embodiment 20. The system of embodiment 19, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • the system of embodiment 19, wherein the lipid nanoparticle is about no greater than about 50 nm.
  • Embodiment 22 The system of any one of the preceding embodiments, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • Embodiment 23 The system of embodiment 22, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 24 The system of embodiment 22, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 25 The system of any one of the preceding embodiments, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • Embodiment 26 The system of any one of the preceding embodiments, wherein the static mixer comprises 8 or more elements.
  • Embodiment 27 The system of any one of the preceding embodiments, wherein the static mixer comprises 8 or more elements.
  • the static mixer comprises 12 or more elements.
  • Embodiment 28 The system of any one of the preceding embodiments, wherein the static mixer comprises 16 or more elements.
  • Embodiment 29 The system of any one of the preceding embodiments, wherein the static mixer comprises 24 or more elements.
  • Embodiment 30 The system of any one of the preceding embodiments, wherein the static mixer is downstream of the conduit.
  • Embodiment 31 A method of generating a lipid nanoparticle, the method comprising: (a) providing: Atty Dkt No.: 58530-742601 i . a conduit comprising: a. a first inlet; b. a second inlet; c.
  • Embodiment 32 The method of embodiment 31, wherein the conduit is tee-shaped.
  • Embodiment 33 The method of embodiment 31, wherein the conduit is Y-shaped.
  • Embodiment 34 The method of embodiment 31, wherein the conduit is Y-shaped.
  • Embodiment 33 wherein the conduit further comprises a cut zip tie.
  • Embodiment 35 The method of embodiment 31, wherein the conduit is L-shaped.
  • Embodiment 36 The method of any one of embodiments 31-35, wherein the static mixer is less than about 40 mm in length.
  • Embodiment 37 The method of any one of embodiments 31-36, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 38 The method of any one of embodiments 31-37, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 39 The method of any one of embodiments 31-38, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 40 The method of any one of embodiments 31-38, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 41 The method of embodiment 40, wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 42 The method of embodiment 40, wherein the inner diameter is about 1/8 inch.
  • Embodiment 43 The method of any one of embodiments 31-42, wherein the polydispersity index is about 0.05 to about 0.25. Atty Dkt No.: 58530-742601 Embodiment 44. The method of embodiment 43, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 45.
  • Embodiment 46 The method of any one of embodiments 31-42, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 46 The method of any one of embodiments 31-45, wherein a Reynolds number of the mixture fluid is about 150 to about 1000.
  • Embodiment 47 The method of embodiment 46, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 48 The method of embodiment 46, wherein the Reynolds number is about 1000.
  • Embodiment 49 The method of any one of embodiments 31-48, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 50 The method of embodiment 49, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 51 The method of any one of embodiments 31-42, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 46 The method of any one of embodiments 31-45, wherein a Reynolds number of the mixture fluid is about 150
  • Embodiment 52 The method of any one of embodiments 31-51, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • Embodiment 53 The method of embodiment 52, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 54 The method of embodiment 52, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 55 Embodiment 55.
  • a system for generating a lipid nanoparticle comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; i ii.
  • Embodiment 62 The system of embodiment 61, wherein the conduit is tee-shaped.
  • Embodiment 63 The system of embodiment 61, wherein the conduit is Y-shaped.
  • Embodiment 64 The system of embodiment 63, wherein the conduit further comprises a cut zip tie.
  • Embodiment 65 The system of embodiment 61, wherein the conduit is L-shaped.
  • Embodiment 61-65 The system of any one of embodiments 61-65, wherein the static mixer is less than about 40 mm in length.
  • Embodiment 67 The system of any one of embodiments 61-66, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 68 The system of any one of embodiments 61-67, wherein the static mixer is less than about 5 mm in length.
  • the system of any one of embodiments 61-68, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 70 The system of any one of embodiments 61-69, wherein the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch.
  • Embodiment 71 The system of any one of embodiments 61-69, wherein the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch.
  • Embodiment 70 wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 72 The system of embodiment 70, wherein the inner diameter is about 1/8 inch.
  • Embodiment 73 The system of any one of embodiments 61-72, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 74 The system of embodiment 73, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 75 The system of embodiment 70, wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 72 The system of embodiment 70, wherein the inner diameter is about 1/8 inch.
  • Embodiment 73 The system of any one of embodiments 61-72, wherein the nucle
  • Embodiment 76 The system of any one of embodiments 61-72, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 76 The system of any one of embodiments 61-75, wherein a Reynolds number of the mixture fluid is about 150 to about 1000.
  • Embodiment 77 The system of embodiment 76, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 78 The system of embodiment 76, wherein the Reynolds number is about 1000.
  • Embodiment 79 The system of any one of embodiments 61-78, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 80 The system of embodiment 79, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 81 The system of embodiment 79, wherein the lipid nanoparticle is about no greater than about 50 nm.
  • Embodiment 82 The system of any one of embodiments 61-81, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • Embodiment 83 The system of embodiment 82, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 84 The system of embodiment 82, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 85 Embodiment 85.
  • Embodiment 90 The system of any one of embodiments 61-89, wherein the static mixer is downstream of the conduit.
  • Embodiment 91 A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit comprising: a. a first inlet; b. a second inlet; c. a junction site downstream of the first inlet and the second inlet; and ii.
  • a static mixer wherein the static mixer is disposed at a distal end of the junction site; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the junction site; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site.
  • Embodiment 92 The method of embodiment 91, wherein the conduit is tee-shaped.
  • Embodiment 93 The method of embodiment 91, wherein the conduit is Y-shaped.
  • Embodiment 94 The method of embodiment 93, wherein the conduit further comprises a cut zip tie.
  • Embodiment 95 The method of embodiment 91, wherein the conduit is L-shaped.
  • Embodiment 96 The method of any one of embodiments 91-95, wherein the static mixer is less than about 40 mm in length.
  • Embodiment 97 The method of any one of embodiments 91-96, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 98 The method of any one of embodiments 91-97, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 99 The method of any one of embodiments 91-98, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 100 The method of any one of embodiments 91-99, wherein the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch.
  • Embodiment 101 The method of embodiment 100, wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 102 The method of embodiment 100, wherein the inner diameter is about 1/8 inch.
  • Embodiment 103 The method of any one of embodiments 91-102, wherein the nucleic acid and the one or more lipid components of I are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 104 The method of any one of embodiments 91-102, wherein the nucleic acid and the one or more lipid components of I are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 105 The method of any one of embodiments 91-102, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 106 The method of any one of embodiments 91-105, wherein a Reynolds number of the mixture fluid is about 150 to about 1000.
  • Embodiment 107 The method of embodiment 106, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 108 The method of embodiment 106, wherein the Reynolds number is about 1000.
  • Embodiment 109 The method of any one of embodiments 91-108, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 110 The method of any one of embodiments 91-108, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 110 The method of any one of embodiments 91-108, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 110 The method of any one of embodiments
  • Embodiment 109 wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 111 The method of embodiment 109, wherein the lipid nanoparticle is about no greater than about 50 nm.
  • Embodiment 112. The method of any one of embodiments 91-111, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • Embodiment 113 The method of embodiment 112, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 114 The method of embodiment 114.
  • Embodiment 112 wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 115 The method of any one of embodiments 91-114, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby Atty Dkt No.: 58530-742601 encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • Embodiment 116 The method of any one of embodiments 91-115, wherein the static mixer comprises 8 or more elements.
  • Embodiment 117 The method of any one of embodiments 91-116, wherein the static mixer comprises 12 or more elements.
  • a system for generating a lipid nanoparticle comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; i ii.
  • Embodiment 122 The system of embodiment 121, wherein the conduit is tee-shaped.
  • Embodiment 123 The system of embodiment 121, wherein the conduit is Y-shaped.
  • Embodiment 124. The system of embodiment 123, wherein the conduit further comprises a cut zip tie.
  • Embodiment 125 The system of embodiment 121, wherein the conduit is L-shaped.
  • Embodiment 129 The system of any one of embodiments 121-125, wherein the static mixer is less than about 40 mm in length.
  • Embodiment 127 The system of any one of embodiments 121-126, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 128 The system of any one of embodiments 121-127, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 129 The system of any one of embodiments 121-128, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 130 The system of any one of embodiments 121-129, wherein the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch.
  • Embodiment 131 The system of embodiment 130, wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 132 The system of embodiment 130, wherein the inner diameter is about 1/8 inch.
  • Embodiment 133 The system of any one of embodiments 121-132, wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 134 The system of embodiment 133, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 135. The system of any one of embodiments 121-132, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 136 The system of any one of embodiments 121-135, wherein the Reynolds number is about 150 to about 1000.
  • Embodiment 137 The system of embodiment 136, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 138 The system of embodiment 136, wherein the Reynolds number is about 1000.
  • Embodiment 139 The system of any one of embodiments 121-138, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 140 The system of embodiment 139, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 141 The system of embodiment 139, wherein the lipid nanoparticle is about no greater than about 50 nm.
  • Embodiment 142 The system of embodiment 136, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 138 The system of embodiment 136, wherein the Reynolds number is about 1000.
  • Embodiment 139 The system of any one of embodiments 121-138, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 140
  • Embodiment 143 The system of embodiment 142, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 144 The system of embodiment 142, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Atty Dkt No.: 58530-742601 Embodiment 145.
  • Embodiment 146 The system of any one of embodiments 121-145, wherein the static mixer comprises 8 or more elements.
  • Embodiment 147 The system of any one of embodiments 121-146, wherein the static mixer comprises 12 or more elements.
  • Embodiment 148 The system of any one of embodiments 121-147, wherein the static mixer comprises 16 or more elements.
  • Embodiment 149 The system of any one of embodiments 121-148, wherein the static mixer comprises 24 or more elements.
  • Embodiment 150 The system of any one of embodiments 121-149, wherein the static mixer is downstream of the conduit.
  • Embodiment 151. A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit comprising: a. a first inlet; b. a second inlet; c. an outlet; and ii.
  • a static mixer (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid, wherein a Reynolds number of the mixture fluid is no greater than about 1000.
  • Embodiment 153. The method of embodiment 151, wherein the conduit is Y-shaped.
  • Embodiment 153 wherein the conduit further comprises a cut zip tie.
  • Embodiment 155 The method of embodiment 151, wherein the conduit is L-shaped. Atty Dkt No.: 58530-742601
  • Embodiment 156 The method of any one of embodiments 151-155, wherein the static mixer is less than about 40 mm in length.
  • Embodiment 157 The method of any one of embodiments 151-156, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 158 The method of any one of embodiments 151-157, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 159 The method of any one of embodiments 151-157, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 160 The method of any one of embodiments 151-158, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 160. The method of any one of embodiments 151-159, wherein the static mixer has an inner diameter of about 1/16 inch to abo1 ⁇ 21/2 inch. Embodiment 161.
  • the method of embodiment 160, wherein the inner diameter is abo1 ⁇ 41/4 inch.
  • Embodiment 162. The method of embodiment 160, wherein the inner diameter is about 1/8 inch.
  • Embodiment 163 The method of any one of embodiments 151-162, wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 164. The method of embodiment 163, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 165 The method of any one of embodiments 151-162, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 166 The method of any one of embodiments 151-165, wherein the Reynolds number is about 150 to about 1000.
  • Embodiment 167 The method of embodiment 166, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 168 The method of embodiment 166, wherein the Reynolds number is about 1000.
  • Embodiment 169 The method of any one of embodiments 151-168, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 170 The method of embodiment 169, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 171 The method of embodiment 169, wherein the lipid nanoparticle is about no greater than about 50 nm. Atty Dkt No.: 58530-742601 Embodiment 172.
  • the method of embodiment 172, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 174 The method of embodiment 172, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 175. The method of any one of embodiments 151-174, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • Embodiment 176. The method of any one of embodiments 151-175, wherein the static mixer comprises 8 or more elements.
  • Embodiment 177. The method of any one of embodiments 151-176, wherein the static mixer comprises 12 or more elements.
  • Embodiment 178. The method of any one of embodiments 151-177, wherein the static mixer comprises 16 or more elements.
  • a system for generating a lipid nanoparticle comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; i ii.
  • Embodiment 182 The system of embodiment 181, wherein the conduit is tee-shaped. Embodiment 183. The system of embodiment 181, wherein the conduit is Y-shaped. Embodiment 184. The system of embodiment 183, wherein the conduit further comprises a cut zip tie. Atty Dkt No.: 58530-742601 Embodiment 185. The system of embodiment 181, wherein the conduit is L-shaped. Embodiment 186.
  • Embodiment 187 The system of any one of embodiments 181-186, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 188 The system of any one of embodiments 181-187, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 189 The system of any one of embodiments 181-188, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 190 The system of any one of embodiments 181-189, wherein the static mixer has an inner diameter of about 1/16 inch to abo1 ⁇ 21/2 inch. Embodiment 191.
  • Embodiment 190 wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 192 The system of embodiment 190, wherein the inner diameter is about 1/8 inch.
  • Embodiment 193. The system of any one of embodiments 181-192, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 194. The system of embodiment 193, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 195 The system of any one of embodiments 181-192, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 196 The system of any one of embodiments 181-192, wherein the polydispersity index is no greater than about 0.1.
  • a Reynolds number of the mixture fluid is about 150 to about 1000.
  • the system of embodiment 196, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 198. The system of embodiment 196, wherein the Reynolds number is about 1000.
  • Embodiment 199. The system of any one of embodiments 181-198, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 200 The system of embodiment 199, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Atty Dkt No.: 58530-742601 Embodiment 201.
  • Embodiment 202 The system of any one of embodiments 181-201, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • Embodiment 203 The system of embodiment 202, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 204 The system of embodiment 202, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 205 The system of embodiment 202, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 211 A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit comprising: a. a first inlet; b. a second inlet; c. an outlet; and ii.
  • a static mixer (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and Atty Dkt No.: 58530-742601 ( e) using the static mixer to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min.
  • Embodiment 212 The method of embodiment 211, wherein the conduit is tee-shaped.
  • Embodiment 213. The method of embodiment 211, wherein the conduit is Y-shaped.
  • Embodiment 214 The method of embodiment 211, wherein the conduit is Y-shaped.
  • the conduit further comprises a cut zip tie.
  • Embodiment 215. The method of embodiment 211, wherein the conduit is L-shaped.
  • Embodiment 216. The method of any one of embodiments 211-215, wherein the static mixer is less than about 40 mm in length.
  • Embodiment 217. The method of any one of embodiments 211-216, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 218 The method of any one of embodiments 211-217, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 219. The method of any one of embodiments 211-218, wherein the static mixer is about 4.8 mm in length.
  • the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch.
  • Embodiment 221. The method of embodiment 220, wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 222 The method of embodiment 220, wherein the inner diameter is about 1/8 inch.
  • Embodiment 223. The method of any one of embodiments 211-222, wherein the nucleic acid and the one or more lipid components of € are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 224. The method of embodiment 223, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 226 The method of any one of embodiments 211-222, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 226 The method of any one of embodiments 211-225, wherein a Reynolds number of the mixture fluid is about 150 to about 1000.
  • Embodiment 227 The method of embodiment 226, wherein the Reynolds number is about 500 to about 1000.
  • the method of embodiment 226, wherein the Reynolds number is about 1000.
  • Embodiment 229. The method of any one of embodiments 211-228, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 230.
  • Embodiment 229 wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 231. The method of embodiment 229, wherein the lipid nanoparticle is about no greater than about 50 nm.
  • Embodiment 232. The method of any one of embodiments 211-231, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • Embodiment 233 The method of embodiment 232, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 235 The method of any one of embodiments 211-234, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • Embodiment 236 The method of any one of embodiments 211-235, wherein the static mixer comprises 8 or more elements.
  • Embodiment 237 The method of any one of embodiments 211-236, wherein the static mixer comprises 12 or more elements.
  • Embodiment 238 The method of any one of embodiments 211-234, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • a system for generating a lipid nanoparticle comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; Atty Dkt No.: 58530-742601 i ii.
  • Embodiment 242 The system of embodiment 241, wherein the conduit is tee-shaped. Embodiment 243. The system of embodiment 241, wherein the conduit is Y-shaped. Embodiment 244. The system of embodiment 243, wherein the conduit further comprises a cut zip tie. Embodiment 245. The system of embodiment 241, wherein the conduit is L-shaped. Embodiment 246.
  • Embodiment 252 The system of embodiment 250, wherein the inner diameter is about 1/8 inch.
  • Embodiment 253. The system of any one of embodiments 241-252, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 254. The system of embodiment 253, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 255 The system of any one of embodiments 241-252, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 256. The system of any one of embodiments 241-255, wherein a Reynolds number of the mixture fluid is about 150 to about 1000.
  • Embodiment 257 The system of embodiment 256, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 258 The system of embodiment 256, wherein the Reynolds number is about 1000.
  • Embodiment 259. The system of any one of embodiments 241-258, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 260. The system of embodiment 259, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • the system of embodiment 259, wherein the lipid nanoparticle is about no greater than about 50 nm.
  • a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • the system of embodiment 262, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • the system of embodiment 262, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 271 A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit comprising: Atty Dkt No.: 58530-742601 a . a first inlet; b. a second inlet; c. an outlet; and ii.
  • Embodiment 272 The method of embodiment 271, wherein the conduit is tee-shaped. Embodiment 273. The method of embodiment 271, wherein the conduit is Y-shaped. Embodiment 274. The method of embodiment 273, wherein the conduit further comprises a cut zip tie. Embodiment 275.
  • Embodiment 271 wherein the conduit is L-shaped.
  • Embodiment 276 The method of any one of embodiments 271-275, wherein the static mixer is less than about 40 mm in length.
  • Embodiment 277 The method of any one of embodiments 271-276, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 278 The method of any one of embodiments 271-277, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 279. The method of any one of embodiments 271-278, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 280 The method of any one of embodiments 271-279, wherein the static mixer has an inner diameter of about 1/16 inch to about 1 ⁇ 2 inch.
  • Embodiment 281. The method of embodiment 280, wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 282. The method of embodiment 280, wherein the inner diameter is about 1/8 inch.
  • Embodiment 283. The method of any one of embodiments 271-282, wherein the nucleic acid and the one or more lipid components of € are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 284. The method of embodiment 283, wherein the polydispersity index is about 0.1 to about 0.2. Atty Dkt No.: 58530-742601 Embodiment 285.
  • Embodiment 286 The method of any one of embodiments 271-282, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 286 The method of any one of embodiments 271-285, wherein a Reynolds number of the mixture fluid is about 150 to about 1000.
  • Embodiment 287 The method of embodiment 286, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 288 The method of embodiment 286, wherein the Reynolds number is about 1000.
  • Embodiment 289. The method of any one of embodiments 271-288, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 290 The method of embodiment 289, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 292 The method of any one of embodiments 271-291, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 293. The method of embodiment 292, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 294. The method of embodiment 292, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 295.
  • Embodiment 300 The method of any one of embodiments 271-299, wherein the static mixer is downstream of the conduit.
  • Embodiment 301. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit, comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; i ii.
  • Embodiment 302. The system of embodiment 301, wherein the conduit is tee-shaped.
  • Embodiment 303 The system of embodiment 301, wherein the conduit is Y-shaped.
  • Embodiment 304 The system of embodiment 303, wherein the conduit further comprises a cut zip tie.
  • Embodiment 305 The system of embodiment 301, wherein the conduit is L-shaped.
  • Embodiment 312 The system of embodiment 310, wherein the inner diameter is about 1/8 inch. Atty Dkt No.: 58530-742601 Embodiment 313.
  • the system of embodiment 313, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 315 The system of any one of embodiments 301-312, wherein the polydispersity index is no greater than about 0.1.
  • a Reynolds number of the mixture fluid is about 150 to about 1000.
  • Embodiment 317 The system of embodiment 316, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 318 The system of embodiment 316, wherein the Reynolds number is about 1000.
  • Embodiment 319 The system of any one of embodiments 301-318, wherein the lipid nanoparticle is no greater than about 100 nm.
  • Embodiment 320 The system of embodiment 319, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 321. The system of embodiment 319, wherein the lipid nanoparticle is about no greater than about 50 nm.
  • a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • the system of embodiment 322, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • the system of embodiment 322, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 325 The system of any one of embodiments 301-324, wherein the static mixer comprises 8 or more elements.
  • the system of any one of embodiments 301-325, wherein the static mixer comprises 12 or more elements.
  • Embodiment 327 The system of any one of embodiments 301-326, wherein the static mixer comprises 16 or more elements.
  • Embodiment 328. The system of any one of embodiments 301-327, wherein the static mixer comprises 24 or more elements. Atty Dkt No.: 58530-742601 Embodiment 329.
  • a method of generating a lipid nanoparticle comprising: (a) providing: i. a conduit, comprising: a. a first inlet; b. a second inlet; c. an outlet; and ii.
  • a static mixer (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle.
  • Embodiment 331 The method of embodiment 330, wherein the conduit is tee-shaped.
  • Embodiment 332. The method of embodiment 330, wherein the conduit is Y-shaped.
  • Embodiment 333 The method of embodiment 330, wherein the conduit is Y-shaped.
  • Embodiment 334 The method of embodiment 330, wherein the conduit is L-shaped.
  • Embodiment 335 The method of any one of embodiments 330-334, wherein the static mixer is less than about 40 mm in length.
  • Embodiment 336 The method of any one of embodiments 330-335, wherein the static mixer is less than about 10 mm in length.
  • Embodiment 337 The method of any one of embodiments 330-336, wherein the static mixer is less than about 5 mm in length.
  • Embodiment 338 The method of any one of embodiments 330-337, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 339 The method of any one of embodiments 330-337, wherein the static mixer is about 4.8 mm in length.
  • Embodiment 342 wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 344 The method of any one of embodiments 330-341, wherein the polydispersity index is no greater than about 0.1.
  • Embodiment 345 The method of any one of embodiments 330-344, wherein a Reynolds number of the mixture fluid is about 150 to about 1000.
  • Embodiment 346 The method of embodiment 345, wherein the Reynolds number is about 500 to about 1000.
  • Embodiment 347 The method of embodiment 345, wherein the Reynolds number is about 1000.
  • Embodiment 348 The method of any one of embodiments 330-347, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 349.
  • the method of embodiment 348, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 350 The method of embodiment 348, wherein the lipid nanoparticle is about no greater than about 50 nm.
  • Embodiment 351. The method of any one of embodiments 330-350, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • the method of embodiment 351, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 351 wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 354. The method of any one of embodiments 330-353, wherein the static mixer comprises 8 or more elements.
  • Embodiment 355. The method of any one of embodiments 330-354, wherein the static mixer comprises 12 or more elements.
  • Embodiment 356. The method of any one of embodiments 330-355, wherein the static mixer comprises 16 or more elements. Atty Dkt No.: 58530-742601 Embodiment 357.
  • the method of any one of embodiments 330-356, wherein the static mixer comprises 24 or more elements.
  • Embodiment 359. A system for generating a lipid nanoparticle, the system comprising a static mixer, wherein when a nucleic acid and one or more lipid components are joined in the static mixer, the system reaches a pressure of no greater than 8 psig.
  • Embodiment 360. The system of embodiment 359, wherein the system further comprises a first inlet configured to introduce a nucleic acid.
  • Embodiment 361. The system of embodiment 359 or 360, wherein the system further comprises a second inlet configured to introduce one or more lipid components of the lipid nanoparticle.
  • Embodiment 366 wherein the inner diameter is about 1 ⁇ 4 inch.
  • Embodiment 368 The system of embodiment 366, wherein the inner diameter is about 1/8 inch.
  • Embodiment 369 The system of any one of embodiments 359-368, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25.
  • Embodiment 370 The system of embodiment 369, wherein the polydispersity index is about 0.1 to about 0.2.
  • Embodiment 371. The system of any one of embodiments 369-370, wherein the polydispersity index is no greater than about 0.1.
  • the system of any one of embodiments 359-374, wherein the lipid nanoparticle is no greater than about 100 nm.
  • the system of embodiment 375, wherein the lipid nanoparticle is about 50 nm to about 100 nm.
  • Embodiment 378 The system of any one of embodiments 359-377, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5.
  • Embodiment 379 The system of embodiment 378, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2.
  • Embodiment 380 The system of embodiment 378, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3.
  • Embodiment 382. The system of any one of embodiments 359-381, wherein the static mixer comprises 8 or more elements.
  • Embodiment 383. The system of any one of embodiments 359-382, wherein the static mixer comprises 12 or more elements.
  • the system of any one of embodiments 359-383, wherein the static mixer comprises 16 or more elements.
  • Embodiment 385 The system of any one of embodiments 359-384, wherein the static mixer comprises 24 or more elements.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)

Abstract

The present disclosure provides systems and methods for generating a lipid nanoparticle. The system may comprise a conduit comprising a first inlet configured to introduce a nucleic acid. The conduit may further comprise a second inlet configured to introduce one or more lipid components of the lipid nanoparticle. The conduit may further comprise an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet. The system may further comprise a static mixer. The nucleic acid and the one or more lipid components may be mixed in the static mixer to generate a mixture fluid. A polydispersity index of the mixture fluid may be no greater than about 0.25.

Description

Atty Dkt No.: 58530-742601 SYSTEMS AND METHODS FOR GENERATING LIPID NANOPARTICLES CROSS-REFERENCE [0001] This application claims benefit of U.S. Provisional Application No. 63/554,054, filed February 15, 2024, and U.S. Provisional Application No.63/737,069, filed December 20, 2024, each of which is incorporated by reference herein in its entirety. BACKGROUND [0002] Therapeutic nucleic acids, such as those that enable gene silencing, expression and editing possess great potential for use as genetic medicines. Lipid nanoparticles (LNPs) may be used for delivery of nucleic acids for treatment or prevention of various diseases and disorders. SUMMARY [0003] Provided herein are systems for generating lipid nanoparticles. In some embodiments, the system comprises a conduit, a static mixer, wherein a nucleic acid and one or more lipid components are mixed to generate a mixture fluid; and a high performance liquid (HPLC) pump. In some embodiments, provided herein is a system for generating a lipid nanoparticle, the system comprising a conduit and a static mixer, wherein a nucleic acid and one or more lipid components are mixed to generate a mixture fluid, wherein when the nucleic acid and the one or more lipid components are joined in the static mixer, the system reaches a pressure of no greater than 8 pounds per square inch gauge (psig). In some embodiments, the conduit comprises a first inlet configured to introduce the nucleic acid. In some embodiments, the conduit comprises a second inlet configured to introduce one or more lipid components of the lipid nanoparticle. In some embodiments, the conduit comprises an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet. In some embodiments, the conduit comprises a third inlet configured to introduce the nucleic acid or one or more lipid components of the lipid nanoparticle. In some embodiments, the conduit comprises a fourth inlet configured to introduce the nucleic acid or the one or more lipid components of the lipid nanoparticle. In some embodiments, the static mixer comprises a plurality of elements, wherein each element of the static mixer makes a 270° twist every 1/8 inch. In some embodiments, the system comprises a channel housing the static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the channel is square shaped and makes a -180° twist every 1/8 inch. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In Atty Dkt No.: 58530-742601 some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 millimeters (mm) in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index of the mixture fluid is about 0.1 to about 0.2. In some embodiments, the polydispersity index of the mixture fluid is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the diameter of the lipid nanoparticle is no greater than about 100 nanometers (nm). In some embodiments, the diameter of the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the diameter of the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. [0004] Aspects disclosed herein are for a system for generating a lipid nanoparticle, the system comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is no greater than about 0.25. In some embodiments, the conduit is tee-shaped. In some embodiments, the Atty Dkt No.: 58530-742601 conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 millimeters (mm) in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the polydispersity index is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nanometers (nm). In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0005] Aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit comprising: a first inlet; a second inlet; an outlet; and a static mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more Atty Dkt No.: 58530-742601 lipid components, thereby generating a mixture fluid, wherein a polydispersity index of the mixture fluid is no greater than about 0.25. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the polydispersity index is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0006] Aspects disclosed herein are for a system for generating a lipid nanoparticle, the system comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second Atty Dkt No.: 58530-742601 inlet configured to introduce one or more lipid components of the lipid nanoparticle; a junction site wherein the first inlet and the second inlet intersects; and a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L- shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance Atty Dkt No.: 58530-742601 liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0007] Aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit comprising: a first inlet; a second inlet; a junction site downstream of the first inlet and the second inlet; and a static mixer, wherein the static mixer is disposed at a distal end of the junction site; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the junction site; and using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y- shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the Atty Dkt No.: 58530-742601 static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0008] Aspects disclosed herein are for a system for generating a lipid nanoparticle, the system comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a Reynolds number of the mixture fluid is no greater than about 1000. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, the Reynolds number is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more Atty Dkt No.: 58530-742601 lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the system reaches a pressure of no more than 8 psig. In some embodiments, the system further comprises a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0009] In some embodiments, provided herein is a method of generating a lipid nanoparticle, the method comprising providing a conduit and a static mixer, providing a nucleic acid and one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate, introducing the nucleic acid to the first inlet, introducing one or more lipid components of the lipid nanoparticle to the second inlet, joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid comprising the lipid nanoparticle. In some embodiments, the conduit comprises a first inlet. In some embodiments, the conduit comprises a second inlet. In some embodiments, the conduit comprises a third inlet. In some embodiments, the conduit comprises a fourth inlet. In some embodiments, the conduit comprises an outlet. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer comprises a plurality of elements, wherein each element of the static mixer makes a 270° twist every 1/8 inch. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index of the mixture fluid is about 0.1 to about 0.2. In some embodiments, the polydispersity index of the mixture fluid is no greater than about 0.1. In some embodiments, the Reynolds number is Atty Dkt No.: 58530-742601 about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the diameter of the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the diameter of the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the diameter of the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. [0010] Aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit comprising: a first inlet; a second inlet; an outlet; and a static mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid, wherein a Reynolds number of the mixture fluid is no greater than about 1000. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about 1/4 inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, the Reynolds number is Atty Dkt No.: 58530-742601 about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0011] Aspects disclosed herein are for a system for generating a lipid nanoparticle, the system comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about ½ inch. In some embodiments, the inner diameter is about ¼ inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the Atty Dkt No.: 58530-742601 mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0012] Aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit comprising: a first inlet; a second inlet; an outlet; and a static mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of Atty Dkt No.: 58530-742601 about 1/16 inch to about ½ inch. In some embodiments, the inner diameter is about ¼ inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0013] Aspects disclosed herein are for a system for generating a lipid nanoparticle, the system comprising: a conduit comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the lipid nanoparticle is no greater than about 150 nm. In some embodiments, the conduit is tee- shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the Atty Dkt No.: 58530-742601 static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. In some embodiments, the inner diameter is about ¼ inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0014] Aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit comprising: a first inlet; a second inlet; an outlet; and a static mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the lipid nanoparticle is no greater than about 150 nm. In some Atty Dkt No.: 58530-742601 embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L- shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about ½ inch. In some embodiments, the inner diameter is about ¼ inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0015] Aspects disclosed herein are for a system for generating a lipid nanoparticle, the system comprising: a conduit, comprising: a first inlet configured to introduce a nucleic acid; a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; an outlet, Atty Dkt No.: 58530-742601 wherein the nucleic acid and the one or more lipid components are joined in the outlet; and a static mixer configured to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about ½ inch. In some embodiments, the inner diameter is about ¼ inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. [0016] Aspects disclosed herein are for a method of generating a lipid nanoparticle, the method comprising: providing: a conduit, comprising: a first inlet; a second inlet; an outlet; and a static Atty Dkt No.: 58530-742601 mixer; introducing a nucleic acid to the first inlet; introducing one or more lipid components of the lipid nanoparticle to the second inlet; joining the nucleic acid and the one or more lipid components at the outlet; and using the static mixer to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. In some embodiments, the conduit is tee-shaped. In some embodiments, the conduit is Y-shaped. In some embodiments, the conduit further comprises a cut zip tie. In some embodiments, the conduit is L-shaped. In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 10 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about ½ inch. In some embodiments, the inner diameter is about ¼ inch. In some embodiments, the inner diameter is about 1/8 inch. In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. In some embodiments, the static mixer is downstream of the conduit. In some embodiments, the method further comprises providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. Atty Dkt No.: 58530-742601 [0017] Another aspect of the present disclosure provides a non-transitory computer readable medium comprising machine executable code that, upon execution by one or more computer processors, implements any of the methods above or elsewhere herein. [0018] Another aspect of the present disclosure provides a system comprising one or more computer processors and computer memory coupled thereto. The computer memory comprises machine executable code that, upon execution by the one or more computer processors, implements any of the methods above or elsewhere herein. [0019] Provided herein is a system for generating a lipid nanoparticle, comprising (i) a first inlet configured to introduce a nucleic acid, (ii) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle, and (iii) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein when a nucleic acid and one or more lipid components are mixed in the static mixer, the system reaches a pressure of no greater than 8 psig. [0020] In some embodiments, the static mixer is less than about 40 mm in length. In some embodiments, the static mixer is less than about 5 mm in length. In some embodiments, the static mixer is about 4.8 mm in length. In some embodiments, the static mixer has an inner diameter of about 1/16 inch to about ½ inch. In some embodiments, the inner diameter is about ¼ inch. In some embodiments, the inner diameter is about 1/8 inch. [0021] In some embodiments, the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. In some embodiments, the polydispersity index is about 0.1 to about 0.2. In some embodiments, the polydispersity index is no greater than about 0.1. [0022] In some embodiments, a Reynolds number of the mixture fluid is about 150 to about 1000. In some embodiments, the Reynolds number is about 500 to about 1000. In some embodiments, the Reynolds number is about 1000. [0023] In some embodiments, the lipid nanoparticle is no greater than about 100 nm. In some embodiments, the lipid nanoparticle is about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle is about no greater than about 50 nm. [0024] In some embodiments, a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. In some embodiments, the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Atty Dkt No.: 58530-742601 [0025] In some embodiments, the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. [0026] In some embodiments, the static mixer comprises 8 or more elements. In some embodiments, the static mixer comprises 12 or more elements. In some embodiments, the static mixer comprises 16 or more elements. In some embodiments, the static mixer comprises 24 or more elements. [0027] In some embodiments, the method comprises diluting the lipid nanoparticle. In some embodiments, a particle size of the lipid nanoparticle is within 10% of the particle size of the lipid nanoparticle before dilution. [0028] In some embodiments, provided herein is a method of generating a lipid nanoparticle, the method comprising: (a) providing a system provided herein, (b) introducing a nucleic acid to the first inlet, (c) introducing one or more lipid components of the lipid nanoparticle to the second outlet, (d) joining the nucleic acid and the one or more lipid components at the outlet, and I using the static mixer to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle, wherein a particle size of the lipid nanoparticle is selected by modifying the Reynolds number. [0029] In some embodiments, the particle size is increased by decreasing the Reynolds number. In some embodiments, the particle size is decreased by increasing the Reynolds number. [0030] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in this art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive. INCORPORATION BY REFERENCE [0031] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict Atty Dkt No.: 58530-742601 the disclosure contained in the specification, the specification is intended to supersede and/or take precedence over any such contradictory material. BRIEF DESCRIPTION OF THE DRAWINGS [0032] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which: [0033] FIGs. 1A-I show schematics of the nine tested static mixing setups. FIG. 1A and FIG. 1B show systems A and B, both using a Koflo 24-element 1/8” ID disposable static mixer with a polypropylene tube, with A using a tee and B using a Y for joining inlets prior to mixing. FIG. 1C shows system C using a Koflo 12-element 1/4” OD mixing element 1/4” ID platinum cured silicone tubing. FIG. 1D shows system D uses the same mixer and Y as B, but with the mixer pushed partway into the Y. FIG. 1E shows system E which uses the same mixer and Y as B, only with some inlet separation provided by a cut zip tie. FIG. 1F shows system F which is the same as B, but with only 12 mixing elements. FIG.1G shows system G which is a custom mixer with three 12-element 1/8” mixer together in a 3/16” ID silicone tube. FIG.1H shows system H is a custom mixer with 16 mixing elements, some of which are pushed into a tee such that the lipid and buffer are joined during mixing. FIG. 1I shows system I is a custom mixer with 12 mixing elements, and with the mixer pushed into the lipid inlet of a Y junction. [0034] FIG. 2 shows images of food dye tests for systems A-C at Reynolds numbers of 25, 50, and 100. [0035] FIGs. 3A-B show particle diameter plotted against Reynolds number for system B run at 1:2 FRR (FIG.3A) and polydispersity index (PDI) for the same samples shown in FIG.3A (FIG.3B). [0036] FIGs. 4A-F shows schematics of the six static mixing setups (systems J-N). All systems were tested at Re of 1000. FIG.4A shows system J which is essentially system H (FIG.1H) but with the inlet streams switched and a higher total flow rate. FIG.4B shows system K which is essentially system H (FIG.1H) but with a higher total flow rate. FIG.4C shows system L which is System J but with a 1:3 FRR and no in-line dilution. FIG.4D shows system M which is system J but with a 1:5 FRR and no in-line dilution. FIG.4E shows System N which is System J Atty Dkt No.: 58530-742601 but with a shorter connection between the two tees and half as many mixing elements. FIG.4F shows System O has similar design as Systems J-M but without mixing elements. [0037] FIGs. 5A-B show bar graphs of post-dilution particle diameter (FIG. 5A) and post- dilution PDI (FIG.5B) for mixing systems Tee, J, K, L, M, N, and O. [0038] FIGs. 6A-B show plots of post-dilution and 15 minutes post-mix particle diameter (FIG. 6A) and PDI (FIG.6B). [0039] FIGs. 7A-C shows bar graphs of post-storage particle diameter (FIG. 7A), post-storage PDI (FIG.7B), and post-storage encapsulation efficiency (FIG.7C) for each of the five tested compositions, also showing a static mixing sample compared to a previously formulated cross- tee sample. Post-storage conditions include post-storage at about 2 – about 8°C and post-storage at about -80°C. [0040] FIGs. 8A-B show a splitter insert designed to split flow prior to mixing elements, ensuring consistent mixing. [0041] FIGs. 9A-C show a mixer (FIGs. 9A, C) with a length of 1 inch (25.4 mm) and a diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.9B), each 1/8 inches (3.175 mm) in length that make a 180° twist. The cross-section thickness if about 0.0394 inches (1mm). [0042] FIGs. 10A-B show a mixer (FIG. 10A) and mixing simulation (FIG. 10B) where the mass fraction of ethanol was simulated using Ansys Fluent. The ethanol flow rate is 75.4 mL/min and the water flow rate is 150.8 mL/min. [0043] FIGs. 11A-C show results from two different viscosity models, k-omega with SST (FIG. 11A) and transition-SST (FIG.11B), which is more rigorous for transition-range Reynolds numbers. Both models appeared similar with respect to mixing estimation, as shown by cross- sections (FIGs.11A-B) and an extrapolated plot of standard deviations of ethanol mass fraction at different cross-sections through the mixer (FIG.11C). [0044] FIGs. 12A-C show results from two different viscosity models, k-omega with SST (FIG. 12A) and transition-SST (FIG.12B). When comparing shear stress, differences between model outputs are more evident (FIG.12C). Across the board, transition-SST model outputs about 40% higher shear stress than k-omega model. [0045] FIGs. 13A-B show comparison of mixing by outlet. FIG. 13A shows the mass fraction of ethanol for outflow. FIG.13B shows mass fraction of ethanol for the pressure-outlet. [0046] FIGs. 14A-B show comparison of shear stress between outlets. FIG. 14A shows shear stress for the outflow and FIG.14B show shear stress for the pressure-outlet. The shear stress between the two outlets is about the same. Atty Dkt No.: 58530-742601 [0047] FIGs. 15A-C show a mixer with 8 mixing elements. FIG. 15A shows a 3D model of the mixer with eight mixing elements with dimensions, each making a 180° twist. FIG.15B shows an example mixing conduit 3D model made with mixing elements rendered in FIG.15A. FIG. 15C shows a 3D model of fluid volume/path of mixing conduit shown in FIG.15B. [0048] FIGs. 16A-B show a 3D rendering of a 0.5 mm bore size cross mixer fluid volume. FIG. 16A shows an isometric view. FIG.16B shows a top view. [0049] FIGs. 17A-C show results comparing an 8-element static mixer to a 0.5 mm cross mixer. FIG.17A shows comparison of standard deviation of ethanol mass fraction between the 8- element static mixer and the 0.5 mm cross mixer. Time point of zero indicates first time of mixing between ethanol and water streams, and each data point represents the standard deviation across a cross-section perpendicular to flow direction. FIG.17B shows comparison of average shear stress between the 8-element static mixer and the 0.5 mm cross mixer. Each data point represents the area-weighted average shear stress across a cross-section perpendicular to flow direction. FIG.17C shows the maximum system pressure for both the 8-element static mixer and the 0.5 mm cross mixer. [0050] FIGs. 18A-C show a mixer (FIG. 18A, C) with a length of 0.8 inches (20.32 mm) and diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.18B) each 0.1 inches (2.54 mm) in length, each element making a 180° twist. The cross-section thickness is about 0.0394 inches (1 mm). [0051] FIGs. 19A-C show a mixer (FIG. 19A, C) with a length of 1.2 inches (30.48 mm) and diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.19B) each 0.15 inches (3.81 mm) in length, where each element makes a 180° twist. The cross-section thickness is about 0.0394 inches (1 mm). [0052] FIGs. 20A-C show a 3-channel mixer (FIG. 20A, C) with a length of 1 inch (25.4 mm) and diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.20B), each element 1/8 inches in length, and each element making a 180° twist. The cross-section thickness is 0.0197 inches (0.5 mm) and is Y shaped. [0053] FIGs. 21A-C show a 4-channel mixer (FIG. 21A, C) with a length of 1 inch (25.4 mm) and diameter of 1/8 inches (3.175 mm) comprising 8 elements (FIG.21B), each element 1/8 inches in length, and each element making a 180° twist. The cross-section thickness is 0.5 mm and is X shaped. [0054] FIGs. 22A-C show a 90° turn mixer (FIG. 22A, C) with a length of ½ inches (12.7 mm) comprising two channels and is 1/8 inches (3.175 mm) in diameter. The mixer comprises 8 Atty Dkt No.: 58530-742601 elements (FIG.22B), each 1/16 inches (1.59 mm) in length and each making a 90° twist. The cross-section thickness is about 1 mm. [0055] FIGs. 23A-C show a 270° turn mixer (FIGs. 23A, C) with a length of 1.5 inches (38.1 mm) comprising two channels and is 1/8 inches (3.175 mm) in diameter. The mixer comprises 8 elements (FIG.23B), each 3/16 inches (4.76 mm) in length and each making a 270° twist. The cross-section thickness is about 1 mm. [0056] FIG. 24 shows an exemplary static mixing setup with a conduit 2401 and a mixer 2402. The conduit can comprise a first inlet 2403, a second inlet 2404, an outlet 2405, and a junction site 2406. The conduit can have an angle 2407 between the first inlet 2403 and the second inlet 2404. The conduit can have an angle 2408 between the first inlet 2403 and the outlet 2405. The conduit can have an angle 2409 between the second inlet 2404 and the outlet 2405. [0057] FIGs. 25A-C show a Koflo analogue design mixer (Design 1, No. A1), with 2 inlets and helical elements and 180° twists (FIG.25A) and a cylindrical channel that houses the elements, and comprise of 1/8 inch barbs that are 120° apart (FIG.25B, C). [0058] FIGs. 26A-D show a design mixer (Design 2, No. D10), with 2 inlets and helical elements and 270° twists (FIG.26A, B) and a square shaped channel that houses the elements (FIG.26C) and comprise -180° twists (FIG.26D). [0059] FIGs. 27A-D show a design mixer (Design 3, No. D7), with 4 inlets and helical elements and 270° twists (FIG.27A, B) and a square shaped channel that houses the elements (FIG.27C) and comprise -180° twists (FIG.27D). [0060] FIGs. 28A-D show a design mixer (Design 5, No. D6), with 4 inlets and helical elements and 180° twists (FIG.28A, B) and a square shaped channel that houses the elements (FIG.28C) and comprise -180° twists (FIG.28D). [0061] FIG. 29A, FIG. 29B, and FIG. 29C show static mixers as described herein. DETAILED DESCRIPTION [0062] Nucleic acids that enable gene silencing, expression, and editing possess great potential for use in genetic medicines for treatment of various indications, including cancer, inherited genetic disorders, and infectious diseases. Efficacious application of nucleic acids for treatment of the abovementioned classes of disorders generally require the use of viral and non-viral delivery methods to facilitate delivery of the nucleic acid to target cells. Lipid nanoparticles (LNPs) represent a potentially efficacious non-viral delivery platform for nucleic acids. Despite progress in this field, recognized herein is a need for improved synthetic approaches for Atty Dkt No.: 58530-742601 generating LNPs comprising nucleic acids. LNP preparation often faces several challenges including particle size and dispersity control. The effectiveness of LNPs can depend on their size and it can be challenging to consistently prepare particles of the size needed for various applications. Furthermore, LNP preparation may also face the issue of low reproducibility due to the complex production processes involved. Cost and scale-up may also inhibit clinical translation of LNPs as preparation of LNPs at scale may be met with reduced homogeneity or modified properties. Currently, LNPs are prepared by various methods including high pressure homogenization, microemulsion, solvent emulsification/evaporation, and ultrasonication. Each of these methods has their own set of advantages and disadvantages, but a method that allows for reproducible, size controlled, and monodisperse LNPs with high nucleic loadings is desirable to enhance clinical translation and viability of LNPs. [0063] The present disclosure provides systems and methods for generation of LNPs. The LNPs may have a small particle size. The LNPs may have a low polydispersity index in a solution. The systems and methods provided herein may be low-cost and may be scaled up efficiently. Methods of generating LNPs as described herein may comprise using a conduit with one or more inlets. The conduit may have one or more outlets. The conduit may have a static mixer. The static mixer may have improved scalability compared to alternative setups (e.g., tee, cross, or microfluidic setups). The improved scalability may be important for commercial scale manufacturing. Each element may allow for production of LNPs with varying ratios of components. The methods provided herein may utilize lower flow and back-pressure and may aid in achieving better control over the physicochemical properties of the resulting drug product. The use of lower flow and lower back-pressure regime could provide a broader selection of pump options and a better control on LNP formation. The use of static mixers provided herein may be a suitable choice for LNP generation. The methods provided herein may allow for reproducible preparation of LNPs with increased control over properties such as size, monodispersity, scalability, and drug loading. Furthermore, the methods provided herein are versatile, allowing for the substitution, addition, or removal of various LNP components (e.g., allowing for 3-component LNPs, 4-component LNPs, 5-component LNPs, or the like) without the need to sacrifice control over the aforementioned properties. Systems and Methods for Generating Lipid Nanoparticles [0064] The present disclosure provides systems and methods for generating lipid nanoparticles. In some embodiments, the lipid nanoparticle encapsulates a nucleic acid. In some embodiments, the nucleic acid may be, for example, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or Atty Dkt No.: 58530-742601 DNA copies of ribonucleic acid (cDNA). The RNA may be, for example, messenger RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), guide RNAs (gRNAs), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), SmY RNA, spliced leader RNA (SL RNA), CRISPR RNA, long noncoding RNA (lncRNA), microRNA (miRNA), or another suitable RNA. The nucleic acid may be RNA. The nucleic acid may be mRNA. The nucleic acid may be DNA. Conduit [0065] Systems as disclosed herein may comprise a conduit (e.g., as shown as 2401 in FIG. 24). In some embodiments, the conduit comprises one or more inlets (e.g., a first inlet, as shown as 2403 in FIG.24, and a second inlet, as shown as 2404 in FIG.24), an outlet, as shown as 2405 in FIG.24, and a junction site, as shown as 2406 in FIG.24. [0066] In some embodiments, the conduit comprises ten or more inlets. In some embodiments, the ten or more inlets intersect at a junction site. In some embodiments, the conduit comprises one to ten (e.g., or more) inlets. In some embodiments, the conduit comprises one to eight inlets. In some embodiments, the conduit comprises one to five inlets. In some embodiments, the conduit comprises one to three inlets. In some embodiments, the conduit comprise one inlet. In some embodiments, the conduit comprises two inlets. In some embodiments, the conduit comprises three inlets. In some embodiments, the conduit comprises four inlets. In some embodiments, the conduit comprises five inlets. In some embodiments, the conduit comprises six inlets. In some embodiments, the conduit comprises seven inlets. In some embodiments, the conduit comprises eight inlets. In some embodiments, the conduit comprises nine inlets. In some embodiments, the conduit comprises ten inlets. [0067] In some embodiments, the conduit may be tee-shaped, as shown in FIG. 1A. In some embodiments, the conduit may be Y-shaped, as shown in FIGs. 1B-1G, and FIG. 1I. In some embodiments, the Y-shaped conduit may further comprise a cut zip tie, as shown in FIG.1E. In some embodiments, the cut zip tie may provide inlet separation. In some embodiments, the cut zip tie may comprise a makeshift flow splitter. In some embodiments, the conduit may be L- shaped, as shown in FIG.1H, and FIGs.4A-4F. [0068] Each of the inlets of the conduit may be used to introduce one or more components of the lipid nanoparticle. In some instances, each inlet is used to introduce a single component of the lipid nanoparticle. In some instances, an inlet is used to introduce at least two (e.g., 3, 4, or 5) components of the lipid nanoparticle. In some embodiments, an inlet is used to introduce a therapeutic agent (e.g., a nucleic acid) and a second (e.g., or second, third, fourth, and/or fifth) inlet is used to introduce the other components of the lipid nanoparticle. Atty Dkt No.: 58530-742601 [0069] The conduit may comprise a first inlet, as shown as 2403 in FIG. 24. The first inlet may be configured to introduce a nucleic acid. The first inlet may be configured to introduce one or more lipid components of the lipid nanoparticle. [0070] The conduit may comprise a second inlet, as shown as 2404 in FIG. 24. The second inlet may be configured to introduce a nucleic acid. The second inlet may be configured to introduce one or more lipid components of the lipid nanoparticle. [0071] The conduit may comprise a third inlet. The third inlet may be configured to introduce a nucleic acid. The third inlet may be configured to introduce one or more lipid components of the lipid nanoparticle. [0072] The conduit may comprise a fourth inlet. The fourth inlet may be configured to introduce a nucleic acid. The fourth inlet may be configured to introduce one or more lipid components of the lipid nanoparticle. [0073] The conduit may comprise an outlet, as shown as 2405 in FIG. 24. The conduit may comprise an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet. [0074] The conduit may comprise a junction site, wherein the first inlet and the second inlet intersects, as shown as 2406 in FIG.24. In some embodiments, the nucleic acid and one or more lipid components may not contact each other prior to entering the junction site. [0075] The conduit may comprise an angle, as shown as 2407 in FIG. 24, between the first inlet and the second inlet. In some embodiments, the angle between the first inlet and the second inlet may be no more than 210° (e.g., no more than 180°, 160°, 140°, 120°, 100°, 90°, 80°, 70°, 60°, 50°, 40°, 30°, 20°, or 10°). In some embodiments, the angle between the first inlet and the second inlet is at least 10° (e.g., at least 15°, 20°, 25°, 30°, 35°, 40°, 45°, 50°, 60°, 70°, 80°, or 90°). In some embodiments, the angle between the first inlet and the second inlet is from about 10° to about 90° (e.g., about 10° to about 60°, about 10° to about 45°, about 20° to about 90°, about 30° to about 90°, or about 20° to about 45°). In some embodiments, the angle between the first inlet and the second inlet is about 10°, about 15°, about 20°, about 25°, about 30°, about 35°, about 40°, about 45°, about 50°, about 55°, about 65°, about 70°, about 75°, about 80°, or about 90°. In some embodiments, the angle is less than 100°. In some embodiments, the angle is less than 90°. In some embodiments, the angle is about 20° to about 100°. In some embodiments, the angle is about 30° to about 80°. [0076] The conduit may comprise an angle, as shown as 2408 in FIG. 24, between the first inlet and the outlet. In some embodiments, the angle between the first inlet and the outlet may be at Atty Dkt No.: 58530-742601 least 10° (e.g., at least 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, or 170°). In some embodiments, the angle between the first inlet and the outlet is at most 180° (e.g., at most 170°, 160°, 150°, 140°, 130°, 120°, 110°, 100°, 90°, 80°, 70°, 60°, 50°, or 40°). In some embodiments, the angle between the first inlet and the outlet is from about 40° to about 180° (e.g., 40° to about 160°, 60° to about 160°, 80° to about 160°, or about 100° to about 150°. In some embodiments, the angle between the first inlet and the second outlet is about 180°, about 170°, about 160°, about 150°, about 140°, about 130°, about 120°, about 110°, about 100°, about 90°, or about 80°. In some embodiments, the angle is at least 90°. In some embodiments, the angle is at least 120°. In some embodiments, the angle is about 100° to about 170°. [0077] The conduit may comprise an angle, as shown as 2409 in FIG. 24, between the second inlet and the outlet. In some embodiments, the angle between the second inlet and the outlet may be at least 10° (e.g., at least 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, or 170°). In some embodiments, the angle between the second inlet and the outlet is at most 180° (e.g., at most 170°, 160°, 150°, 140°, 130°, 120°, 110°, 100°, 90°, 80°, 70°, 60°, 50°, or 40°). In some embodiments, the angle between the second inlet and the outlet is from about 40° to about 180° (e.g., 40° to about 160°, 60° to about 160°, 80° to about 160°, or about 100° to about 150°. In some embodiments, the angle between the second inlet and the second outlet is about 180°, about 170°, about 160°, about 150°, about 140°, about 130°, about 120°, about 110°, about 100°, about 90°, or about 80°. In some embodiments, the angle is at least 90°. [0078] In some embodiments, the angle between the first inlet and the outlet, and the angle between the second inlet and the outlet are the same. In some embodiments, the angle between the first inlet and the outlet, and the second inlet and the outlet are different. [0079] In some embodiments, the angle between an inlet (e.g., such as a first inlet, second inlet, third inlet, fourth inlet, fifth inlet, etc.) and the outlet may be at least 10° (e.g., at least 20°, 30°, 40°, 50°, 60°, 70°, 80°, 90°, 100°, 110°, 120°, 130°, 140°, 150°, 160°, or 170°). In some embodiments, the angle between an inlet (e.g., such as a first inlet, second inlet, third inlet, fourth inlet, fifth inlet, etc.) and the outlet is at most 180° (e.g., at most 170°, 160°, 150°, 140°, 130°, 120°, 110°, 100°, 90°, 80°, 70°, 60°, 50°, or 40°). In some embodiments, the angle between an inlet (e.g., such as a first inlet, second inlet, third inlet, fourth inlet, fifth inlet, etc.) and the outlet is from about 40° to about 180° (e.g., 40° to about 160°, 60° to about 160°, 80° to about 160°, or about 100° to about 150°. In some embodiments, the angle between an inlet (e.g., such as a first inlet, second inlet, third inlet, fourth inlet, fifth inlet, etc.) and the second outlet is about Atty Dkt No.: 58530-742601 180°, about 170°, about 160°, about 150°, about 140°, about 130°, about 120°, about 110°, about 100°, about 90°, or about 80°. [0080] In some embodiments, a flow rate of the lipid component into any of the inlets herein (e.g., a first inlet, second inlet, third inlet, fourth inlet, etc.) may be no greater than about 500 mL/min, 300 mL/min, 200 mL/min, 150 mL/min, 100 mL/min, 50 mL/min, 40 mL/min, 30 mL/min, 20 mL/min, 10 mL/min, or 5 mL/min. In some embodiments, a flow rate of the lipid component into any of the inlets herein (e.g., a first inlet, second inlet, third inlet, fourth inlet, etc.) is at least 5 mL/min, 10 mL/min, 20 mL/min, 30 mL/min, 40 mL/min, 50 mL/min, 60 mL/min, or 70 mL/min. In some embodiments, a flow rate of the lipid component any of the inlets herein (e.g., a first inlet, second inlet, third inlet, fourth inlet, etc.) is from about 5 mL/min to about 100 mL/min, about 5 mL/min to about 80 mL/min, about 10 mL/min to about 80 mL/min, about 20 mL/min to about 80 mL/min, about 30 mL/min to about 80 mL/min, about 40 mL/min to about 80 mL/min, or about 50 mL/min to about 80 mL/min. In some embodiments, a flow rate of the lipid component into any of the inlets herein (e.g., a first inlet, second inlet, third inlet, fourth inlet, etc.) is from about 50 mL/min to about 80 mL/min. Increased flow rates may be useful to allow adoption into larger scale production. [0081] In some embodiments, a flow rate of the nucleic acid into the first inlet may be no greater than about 1200 milliliters per minute (mL/min), 1100 mL/min, 1000 mL/min, 900 mL/min, 800 mL/min, 700 mL/min,500 mL/min, 300 mL/min, 200 mL/min, 150 mL/min, 100 mL/min, 50 mL/min, 40 mL/min, 30 mL/min, 20 mL/min, 10 mL/min, or 5 mL/min. In some embodiments, a flow rate of the nucleic acid into the first inlet may be at least 1 mL/min, 5 mL/min, 10 mL/min, 20 mL/min, 30 mL/min, 40 mL/min, 50 mL/min, or 100 mL/min. In some embodiments, a flow rate of the nucleic acid into the first inlet may be from about 5 mL/min to about 300 mL/min, from about 5 mL/min to about 200 mL/min, from about 20 mL/min to about 200 mL/min, from about 40 mL/min to about 200 mL/min, from about 50 mL/min to about 200 mL/min, from about 100 mL/min to about 200 mL/min, or from about 120 mL/min to about 190 mL/min. In some embodiments, the flow rate of the nucleic acid into the first inlet is about 10 mL/min, such as described in Table 5. In some embodiments, the flow rate of the nucleic acid into the first inlet is about 150 mL/min to about 160 mL/min, such as described in Table 5. In some embodiments, the flow rate of the nucleic acid into the first inlet is about 150 mL/min, such a described in Table 5 or Table 7. In some embodiments, the flow rate of the nucleic acid into the first inlet is from about 200 mL/min to about 700 mL/min. Atty Dkt No.: 58530-742601 [0082] In some embodiments, a flow rate of the lipid component into the second inlet may be no greater than about 1200 mL/min, 1100 mL/min, 1000 mL/min, 900 mL/min, 800 mL/min, 700 mL/min, 500 mL/min, 300 mL/min, 200 mL/min, 150 mL/min, 100 mL/min, 50 mL/min, 40 mL/min, 30 mL/min, 20 mL/min, 10 mL/min, or 5 mL/min. In some embodiments, a flow rate of the lipid component into the second inlet is at least 5 mL/min, 10 mL/min, 20 mL/min, 30 mL/min, 40 mL/min, 50 mL/min, 60 mL/min, 70 mL/min, 100 mL/min, 250 mL/min, 300 mL/min, 400 mL/min, 500 mL/min, 600 mL/min, 700 mL/min, or 800 mL/min. In some embodiments, a flow rate of the lipid component into the second inlet is from about 5 mL/min to about 1200 mL/min, about 100 mL/min to about 1000 mL/min, about 5 mL/min to about 100 mL/min, about 5 mL/min to about 80 mL/min, about 10 mL/min to about 80 mL/min, about 20 mL/min to about 80 mL/min, about 30 mL/min to about 80 mL/min, about 40 mL/min to about 80 mL/min, or about 50 mL/min to about 80 mL/min. In some embodiments, a flow rate of the lipid component into the second inlet is from about 50 mL/min to about 80 mL/min. In some embodiments, a flow rate of the lipid component into the second inlet is about 75 mL/min, such as described in Table 5 and Table 7. In some embodiments, a flow rate of the lipid component into the second inlet is about 55 mL/min, such as described in Table 5. In some embodiments, a flow rate of the lipid component into the second inlet is about 5, such as described in Table 5. In some embodiments, a flow rate of the lipid component into the second inlet is from about 50 mL/min to about 300 mL/min. [0083] In some embodiments, a ratio between a flow rate of the lipid component into any two of the inlets provided herein (e.g., a first, second, third, fourth, fifth, etc.) is from about 1:1 to about 1:10, about 1:1 to about 1:8, about 1:1 to about 1:7, about 1:1 to about 1:6, about 1:1 to about 1:3 or about 1:1 to about 1:4. In some embodiments, a ratio between a flow rate of the lipid component into any two of the inlets provided herein (e.g., a first, second, third, fourth, fifth, etc.) is at least about 1:1, 1:1.25, 1:1.5, 1:1.75, 1:2, 1:2.5, 1:3, or 1:4. In some embodiments, a ratio between a flow rate of the lipid component into any two of the inlets provided herein (e.g., a first, second, third, fourth, fifth, etc.) is at most about 1:10, 1:8, 1:6, 1:5, 1:4, 1:3, or 1:2. In some embodiments, a ratio between a flow rate of the lipid component into any two of the inlets provided herein (e.g., a first, second, third, fourth, fifth, etc.) may be about 1:2, about 1:3, about 1:5, about 1:8, or about 1:10. [0084] In some embodiments, a ratio between a flow rate of the lipid component into the second inlet and a flow rate of the nucleic acid into the first inlet is from about 1:1 to about 1:10, about 1:1 to about 1:8, about 1:1 to about 1:7, about 1:1 to about 1:6, about 1:1 to about 1:3 or about Atty Dkt No.: 58530-742601 1:1 to about 1:4. In some embodiments, a ratio between a flow rate of the lipid component into the second inlet and a flow rate of the nucleic acid into the first inlet is at least about 1:1, 1:1.25, 1:1.5, 1:1.75, 1:2, 1:2.5, 1:3, or 1:4. In some embodiments, a ratio between a flow rate of the lipid component into the second inlet and a flow rate of the nucleic acid into the first inlet is at most about 1:10, 1:8, 1:6, 1:5, 1:4, 1:3, or 1:2. In some embodiments, a ratio between a flow rate of the lipid component into the second inlet and a flow rate of the nucleic acid into the first inlet may be about 1:2, about 1:3, about 1:5, about 1:8, or about 1:10. Inlet to Introduce Nucleic Acid [0085] In some embodiments, the first inlet may be configured to introduce a therapeutic agent, such as a nucleic acid. In some embodiments, the first inlet may be configured to introduce a nucleic acid. In some embodiments, the second inlet may be configured to introduce a therapeutic agent, such as a nucleic acid. In some embodiments, the second inlet may be configured to introduce a nucleic acid. In some embodiments, the third inlet may be configured to introduce a therapeutic agent, such as a nucleic acid. In some embodiments, the third inlet may be configured to introduce a nucleic acid. In some embodiments, the fourth inlet may be configured to introduce a therapeutic agent, such as a nucleic acid. In some embodiments, the fourth inlet may be configured to introduce a nucleic acid. In some embodiments, the nucleic acid may comprise a small interfering ribonucleic acid (siRNA), a short hairpin RNA (shRNA), a micro-ribonucleic acid (miRNA), a primary micro-ribonucleic acid (pri-miRNA), a long non- coding RNA (lncRNA), a messenger ribonucleic acid (mRNA), a clustered regularly interspaced short palindromic repeats (CRISPR) related nucleic acid, a CRISPR-RNA (crRNA), a single guide ribonucleic acid (sgRNA), a trans-activating CRISPR ribonucleic acid (tracrRNA), a plasmid deoxyribonucleic acid (pDNA), a transfer ribonucleic acid (tRNA), an antisense oligonucleotide (ASO), an antisense ribonucleic acid (RNA), a guide ribonucleic acid, deoxyribonucleic acid (DNA), a double stranded deoxyribonucleic acid (dsDNA), a single stranded deoxyribonucleic acid (ssDNA), a single stranded ribonucleic acid (ssRNA), a or double stranded ribonucleic acid (dsRNA). In some embodiments, the first inlet may comprise a nucleic acid. In some embodiments, the first inlet may comprise mRNA. In some embodiments, the second inlet may comprise a nucleic acid. In some embodiments, the second inlet may comprise mRNA. In some embodiments, the third inlet may comprise a nucleic acid. In some embodiments, the third inlet may comprise mRNA. In some embodiments, the fourth inlet may comprise a nucleic acid. In some embodiments, the fourth inlet may comprise mRNA. In some Atty Dkt No.: 58530-742601 embodiments, the mRNA is at least 15 mg (e.g., at least 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 80 mg, or 100 mg). [0086] In some embodiments, the lipid nanoparticle comprises one or more nucleic acids present in a weight ratio to the cationic ionizable lipid of from about 5:1 to about 1:100. In some embodiments, the weight ratio of the one or more nucleic acids to cationic ionizable lipid is at least 2:1, 1:1, 1:2, 1:5, 1:10, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, or 1:50. In some embodiments, the weight ratio of the one or more nucleic acids to cationic ionizable lipids is at most 1:100, 1:90, 1:80, 1:70, 1:60, 1:50, 1:45, 1:40, 1:35, or 1:30. In some embodiments, the weight ratio of nucleic acid to cationic ionizable lipid is from about 5:1, 2.5:1, 1:1, 1:5, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:60, 1:70, 1:80, 1:90, or 1:100, or any range derivable therein. In some embodiments, the weight ratio is about 1:25 or about 1:7. In some embodiments, the weight ratio is from about 1:30 to about 1:40. In some embodiments, the weight ratio is about 1:30, 1:35, or about 1:40. Inlet to Introduce Lipid Components [0087] In some embodiments, any one of the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth inlet may be configured to introduce one or more lipid components of the lipid nanoparticle. A lipid nanoparticle may comprise 2, 3, 4, 5, 6 or more components. In some embodiments, a lipid nanoparticle comprises 2 components. In some embodiments, a lipid nanoparticle comprises 3 components. In some embodiments, a lipid nanoparticle comprises 4 components. In some embodiments, a lipid nanoparticle comprises 5 components. In some embodiments, a lipid nanoparticle comprises 6 (e.g., or more) components. In some embodiments, a lipid nanoparticle comprises one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, and a PEGylated lipid. In some embodiments, a lipid nanoparticle comprises a nucleic acid and one or more lipid components (e.g., a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, and a PEGylated lipid). In some embodiments, the first inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the second inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the third inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the fourth inlet may be configured to introduce one or Atty Dkt No.: 58530-742601 more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the fifth inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the sixth inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the seventh inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the eighth inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the ninth inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. In some embodiment, the tenth inlet may be configured to introduce one or more of a therapeutic agent (e.g., a nucleic acid), a cationic ionizable lipid, a SORT lipid, a phospholipid, a steroid, or a PEGylated lipid. [0088] In some instances, the lipid nanoparticles provided herein comprise a cationic ionizable lipid. Cationic ionizable lipids as described elsewhere in the field may be used herein. For instance, examples of cationic ionizable lipids may be found at U.S. Patent No.11,542,229, U.S. Patent No.11,247,968, U.S. Patent No.11,766,408, and U.S. Patent No.11,229,609, each of which are incorporated by reference herein in their entireties. [0089] In some embodiments, the cationic ionizable lipid is present in a molar percentage of the total lipid components of from about 2% to about 60%. In some embodiments, the molar percentage of the cationic ionizable lipid is from about 5% to about 50%. In some embodiments, the molar percentage of the cationic ionizable lipid is from about 5% to about 30%. In some embodiments, the molar percentage of the cationic ionizable lipid is from about 7.5% to about 20%. In some embodiments, the molar percentage of the cationic ionizable lipid is at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the molar percentage of the cationic ionizable lipid is at most 70%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, or 25%. [0090] The lipid nanoparticles may comprise a steroid or steroid derivative. In some embodiments, the steroid or steroid derivative comprises any steroid or steroid derivative. As used herein, in some embodiments, the term “steroid” is a class of compounds with a four ring Atty Dkt No.: 58530-742601 17 carbon cyclic structure which can further comprises one or more substitutions including alkyl groups, alkoxy groups, hydroxy groups, oxo groups, acyl groups, or a double bond between two or more carbon atoms. Examples of steroids may be found at U.S. Patent No.11,542,229, U.S. Patent No.11,247,968, U.S. Patent No.11,766,408, and U.S. Patent No.11,229,609, each of which are incorporated by reference herein in their entireties. [0091] In some embodiments, the steroid or steroid derivative is present in a molar percentage of the total lipid components of from about 20% to about 60%. In some embodiments, the steroid or steroid derivative is present in a molar percentage of from about 30% to 50%. In some embodiments, the steroid or steroid derivative is present in a molar percentage of from about 39% to about 46%. In some embodiments, the steroid or steroid derivative is present in a molar percentage of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, or 55%. In some embodiments, the steroid or steroid derivative is present in a molar percentage of at most 60%, 55%, 50%, 45%, 40%, 35%, 30%, or 25%. [0092] In some embodiments, the lipid nanoparticles comprise PEG or a PEGylated lipid. In some embodiments, the present disclosure comprises using any lipid to which a PEG group has been attached. Examples of PEG or a PEGylated lipids may be found at U.S. Patent No. 11,542,229, U.S. Patent No.11,247,968, U.S. Patent No.11,766,408, and U.S. Patent No. 11,229,609, each of which are incorporated by reference herein in their entireties. [0093] In some embodiments, the PEG or PEGylated lipid is present in a molar percentage of the total lipid components of from about 0.1% to about 20%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of from about 0.5% to about 10%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of from about 0.5% to about 5%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of from about 2% to about 2.8%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of at least 0.1%, 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, or 8%. In some embodiments, the PEG or PEGylated lipid is present at a molar percentage of at most 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, or 2%. [0094] In some embodiments, the lipid nanoparticles comprise a phospholipid. In some aspects of the present disclosure, the polymers are mixed with one or more phospholipids to create a composition. In some embodiments, any lipid which also comprises a phosphate group. In some embodiments, the phospholipid is a structure which contains one or two long chain C6-C24 alkyl or alkenyl groups, a glycerol or a sphingosine, one or two phosphate groups, and, optionally, a small organic molecule. Examples of phospholipids may be found at U.S. Patent No.11,542,229, Atty Dkt No.: 58530-742601 U.S. Patent No.11,247,968, U.S. Patent No.11,766,408, and U.S. Patent No.11,229,609, each of which are incorporated by reference herein in their entireties. [0095] In some embodiments, the phospholipid is present in a molar percentage of total lipid components of about 1% to about 40%. In some embodiments, the phospholipid is present in a molar percentage of total lipid components of about 10% to about 40%. In some embodiments, the phospholipid is present in a molar percentage of total lipid components of at least about 5%, 7.5%, 10%, 15%, 17%, 18%, 20%, 22%, or 25%. In some embodiments, the phospholipid is present in a molar percentage of total lipid components of at most about 40%, 38%, 35%, 33%, 30%, 28%, 25%, 22%, or 20%. [0096] In some embodiments, the lipid nanoparticles comprise one or more selective organ targeting (SORT) compounds, which lead to the selective delivery of the lipid nanoparticle to a particular organ. This compound may be a lipid, a small molecule therapeutic agent, a sugar, a vitamin, or a protein. Examples of SORT lipids can be found at U.S. Patent No.11,766,408 and U.S. Patent No.11,229,609, which are incorporated by reference herein in their entireties. [0097] In some embodiments, the one or more SORT compounds are present in a molar percentage of total lipid components of from about 5% to about 50%. In some embodiments, the one or more SORT compounds are present in molar percentage of from about 10% to about 45%. In some embodiments, the one or more SORT compounds are present in an amount of about 20% to about 40%. In some embodiments, the one or more SORT compounds are present in an amount of at least 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 50%. In some embodiments, the one or more SORT compounds are present in an amount of at most 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10%. [0098] In some aspects, the present disclosure provides one or more lipids with one or more hydrophobic components and a permanently cationic group. The permanently cationic lipid may contain a group which has a positive charge regardless of the pH. Permanently cationic lipids as described elsewhere in the field may be used. In some instances, examples of permanently cationic groups can be found at U.S. Patent No.11,766,408 and U.S. Patent No.11,229,609, which are incorporated by reference herein in their entireties. [0099] In some embodiments, the lipid nanoparticle may comprise a permanently cationic lipid at any suitable amount. In some instances, examples of permanently cationic lipids can be found at U.S. Patent No.11,766,408 and U.S. Patent No.11,229,609, which are incorporated by reference herein in their entireties. Atty Dkt No.: 58530-742601 [00100] In some aspects, the present disclosure provides one or more lipids with one or more hydrophobic components and a permanently anionic group. In some instances, examples of permanently anionic lipids can be found at U.S. Patent No.11,766,408 and U.S. Patent No. 11,229,609, which are incorporated by reference herein in their entireties. Static Mixer [00101] The present disclosure provides systems comprising a static mixer. In some embodiments, the static mixer may be downstream of the conduit. In some embodiments, the static mixer may be upstream of the conduit. In some embodiments, the static mixer may be parallel to the conduit. [00102] The system may comprise a static mixer. The system may comprise a static mixer, wherein the nucleic acid and one or more lipid components are mixed to generate a mixture fluid, and wherein the nucleic acid and the one or more lipid components may not contact each other prior to entering the junction site. In some embodiments, the nucleic acid and one or more lipid components are mixed to generate a mixture fluid within the static mixer. [00103] A static mixer described herein may comprise a design as illustrated in FIG. 29A, FIG.29B, or FIG.29C. A static mixer described herein may comprise a design as illustrated in FIG.29A, FIG.29B, FIG.29C, or a combination thereof. A static mixer described herein may comprise a design as illustrated in FIG.29A. A static mixer described herein may comprise a design as illustrated in FIG.29B. A static mixer described herein may comprise a design as illustrated in FIG.29C. [00104] A static mixer described herein may comprise design A1, as illustrated in FIG. 29A. A static mixer described herein may comprise design A2, as illustrated in FIG.29A. A static mixer described herein may comprise design A3, as illustrated in FIG.29A. A static mixer described herein may comprise design A4, as illustrated in FIG.29A. A static mixer described herein may comprise design A5, as illustrated in FIG.29A. A static mixer described herein may comprise design A6, as illustrated in FIG.29A. A static mixer described herein may comprise design B1, as illustrated in FIG.29A. A static mixer described herein may comprise design B2, as illustrated in FIG.29A. A static mixer described herein may comprise design B3, as illustrated in FIG.29A. A static mixer described herein may comprise design B4, as illustrated in FIG.29A. A static mixer described herein may comprise design C1, as illustrated in FIG. 29A. A static mixer described herein may comprise design C2, as illustrated in FIG.29A. A static mixer described herein may comprise design C3, as illustrated in FIG.29A. A static mixer described herein may comprise design C4, as illustrated in FIG.29B. A static mixer described Atty Dkt No.: 58530-742601 herein may comprise design C5, as illustrated in FIG.29B. A static mixer described herein may comprise design C6, as illustrated in FIG.29B. A static mixer described herein may comprise design C7, as illustrated in FIG.29B. A static mixer described herein may comprise design C8, as illustrated in FIG.29B. A static mixer described herein may comprise design C9, as illustrated in FIG.29B. A static mixer described herein may comprise design C10, as illustrated in FIG.29B. A static mixer described herein may comprise design C11, as illustrated in FIG. 29B. A static mixer described herein may comprise design C12, as illustrated in FIG.29B. A static mixer described herein may comprise design C13, as illustrated in FIG.29B. A static mixer described herein may comprise design C14, as illustrated in FIG.29B. A static mixer described herein may comprise design C15, as illustrated in FIG.29B. A static mixer described herein may comprise design C16, as illustrated in FIG.29B. A static mixer described herein may comprise design C17, as illustrated in FIG.29C. A static mixer described herein may comprise design C18, as illustrated in FIG.29C. A static mixer described herein may comprise design C18, as illustrated in FIG.29C. A static mixer described herein may comprise design D1, as illustrated in FIG.29C. A static mixer described herein may comprise design D2, as illustrated in FIG.29C. A static mixer described herein may comprise design D3, as illustrated in FIG. 29C. A static mixer described herein may comprise design D4, as illustrated in FIG.29C. A static mixer described herein may comprise design D5, as illustrated in FIG.29C. A static mixer described herein may comprise design D6, as illustrated in FIG.29C. A static mixer described herein may comprise design D7, as illustrated in FIG.29C. A static mixer described herein may comprise design D8, as illustrated in FIG.29C. A static mixer described herein may comprise design D9, as illustrated in FIG.29C. A static mixer described herein may comprise design D10, as illustrated in FIG.29C. [00105] In some embodiments, a static mixer described herein may comprise a design of A1, A2, A3, A4, A5, A6, B1, B2, B3, B4, C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 C11, C12, C13, C14, C15, C16, C17, C18, D1, D2, D3, D4, D5, D6, D7, D8, D9, D10, or any combination thereof. [00106] In some embodiments, the static mixer may have a length less than about 1000 millimeter (mm), 500 mm, 300 mm, 200 mm, 100 mm, 90 mm, 80 mm, 70 mm, 60 mm, 50 mm, 45 mm, 40 mm, 35 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 9 mm, 8 mm, 7 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, or 1 mm. In some embodiments, the static mixer comprises a length of at least 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 15 mm, 20 mm, 25 mm, or 30 mm. In some embodiments, the static mixer comprises a length of from about 5 mm to about 80 mm. In some Atty Dkt No.: 58530-742601 embodiments, the static mixer comprises a length of from about 5 mm to about 40 mm. In some embodiments, the static mixer comprises a length of from about 10 mm to about 35 mm. In some embodiments, the static mixer comprises a length of about 20 mm, as shown in FIGs.18A-18C. In some embodiments, the static mixer comprises a length of about 30 mm, as shown in FIGs. 19A-19C. In some embodiments, the static mixer comprises a length of about 25 mm, as shown in FIGs.20A-20C. In some embodiments, the static mixer comprises a length of about 12 mm, as shown in FIGs.22A-C. In some embodiments, the static mixer may have a length less than about 10 inches (in), 5 in, 4 in, 3 in, 2 in, 1.5 in, 1.2 in, 1 in, 0.9 in, 0.8 in, 0.7 in, 0.6 in, 0.5 in, 0.4 in, 0.3 in, 0.2 in, or 0.1 in. In some embodiments, the static mixer has a length of about 4.8 mm. In some embodiments, the static mixer may have a length about 0.8 in, as shown in FIGs. 18A-18C. In some embodiments, the static mixer may have a length about 1.2 in, as shown in FIGs.19A-19C. In some embodiments, the static mixer may have a length of about 1 in. [00107] In some embodiments, the static mixer may have an inner diameter of less than about 5 in, 4 in, 3 in, 2 in, 1 in, ¾ in, ½ in, 3/8 in, ¼ in, 3/16 in, 1/8 in, 1/16 in, or 1/32 in. In some embodiments, the static mixer comprises an inner diameter of less than about 127 mm, 102 mm, 76 mm, 50 mm, 25 mm, 19 mm, 13 mm, 9.5 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, 1.5 mm, 1 mm, or 0.75 mm. In some embodiments, the static mixer comprises an inner diameter of at least 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 4 mm, or 5 mm. In some embodiments, the static mixer comprises an inner diameter of from about 0.5 mm to about 20 mm. In some embodiments, the static mixer comprises an inner diameter of from about 1 mm to about 10 mm. In some embodiments, the static mixer comprises an inner diameter of from about 1 mm to about 5 mm. In some embodiments, the static mixer comprises an inner diameter of about 3 mm. [00108] In some embodiments, the static mixer may have 1 element, 2 or more elements, 4 or more elements, 6 or more elements, 8 or more elements, 12 or more elements, 16 or more elements, 24 or more elements, or 36 or more elements. In some embodiments, the static mixer comprises at least 1 element, at least 2 elements, at least 3 elements, at least 5 elements, at least 8 elements, at least 12 elements, or at least 20 elements. In some embodiments, the static mixer comprises at most 54 elements, at most 48 elements, at most 36 elements, at most 24 elements, at most 16 elements, or at most 12 elements. In some embodiments, the static mixer comprises from 1 to 12 elements. In some embodiments, the static mixer comprises from 5 to 10 elements. In some embodiments, the static mixer comprises 8 elements. [00109] In some embodiments, the systems provided herein further comprise a splitter insert, which may be arranged within the system in various ways. In some embodiments, the static Atty Dkt No.: 58530-742601 mixer is inserted into the splitter insert, as shown in FIG.8A and FIG.8B. In some embodiments, [00110] The elements of the static mixer may be arranged in various ways. In some embodiments, a splitter insert may be inserted in the static mixer to split the flows prior to mixing the elements and to ensure consistent mixing, as shown in FIG.8A and 8B. In some embodiment, the splitter insert may have a length of less than about 70 mm, 50 mm, 40 mm, 35 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 5 mm, or 1 mm. In some embodiments, the splitter insert comprises a length of at least 1 mm, 5 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 35 mm, or 40 mm. In some embodiments, the splitter insert comprises a length of from about 1 mm to about 50 mm, from about 10 mm to about 40 mm, from about 20 mm to about 40 mm. In some embodiments, the splitter insert comprises a length of from about 30 mm to about 40 mm. In some embodiments, the splitter insert comprises a length of about 35 mm, as shown in FIG. 8A and FIG.8B. In some embodiments, the static mixer comprises a splitter insert of FIG.8A or FIG.8B. [00111] In some embodiments, the splitter insert is a tee-shaped conduit. In other embodiments, the splitter insert is a Y-shaped conduit. In further embodiments, the splitter insert is an L-shaped conduit. [00112] In some embodiments, any suitable number of elements in a static mixer may be inserted into a splitter insert, based on for example, the length of the elements and the length of the splitter insert. In some embodiments, at least 1 of the elements of the static mixer is inserted into the splitter insert. In some embodiments, at least 2, 3, 4, 5, 6, 7, or 8 of the elements of the static mixer may be inserted into the splitter insert. In some embodiments, at most 75% of the elements of the static mixer may be inserted into the splitter insert. In some embodiments, at most 50% of the elements of the static mixer may be inserted into the splitter insert. In some embodiments, at most 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 of the elements of the static mixer may be inserted into the splitter insert. [00113] In some embodiments, any suitable number of elements in a static mixer may be inserted into a tee-shaped conduit, based on for example, the length of the elements and the length of the tee-shaped conduit. In some embodiments, at least 1 of the elements of the static mixer is inserted into the tee-shaped conduit. In some embodiments, at least 2, 3, 4, 5, 6, 7, or 8 of the elements of the static mixer may be inserted into the tee-shaped conduit. In some embodiments, at most 75% of the elements of the static mixer may be inserted into the tee- shaped conduit. In some embodiments, at most 50% of the elements of the static mixer may be Atty Dkt No.: 58530-742601 inserted into the tee-shaped conduit. In some embodiments, at most 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 of the elements of the static mixer may be inserted into the tee-shaped conduit. [00114] In some embodiments, 2 of the 8 or more elements in a static mixer may be inserted into a tee-shaped conduit prior to the static mixer. In some embodiments, 6 of the 8 or more elements in a static mixer may be downstream from an outlet in a tee-shaped conduit. In some embodiments, 4 of the 12 or more elements in a static mixer may be inserted into a tee-shaped conduit prior to the static mixer. In some embodiments, 8 of the 12 or more elements in a static mixer may be downstream from an outlet in the tee-shaped conduit. In some embodiments, 2 of the 16 or more elements in a static mixer may be inserted into a tee-shaped conduit prior to the static mixer. In some embodiments, 4 of the 16 or more elements in a static mixer may be inserted into a tee-shaped conduit prior to the static mixer. In some embodiments, 12 of the 16 or more elements in a static mixer may be downstream from an outlet in the tee-shaped conduit. In some embodiments, 14 of the 16 or more elements in a static mixer may be downstream from an outlet in the tee-shaped conduit. [00115] In some embodiments, any suitable number of elements in a static mixer may be inserted into a Y-shaped conduit, based on for example, the length of the elements and the length of the Y-shaped conduit. In some embodiments, at least 1 of the elements of the static mixer is inserted into the Y-shaped conduit. In some embodiments, at least 2, 3, 4, 5, 6, 7, or 8 of the elements of the static mixer may be inserted into the Y-shaped conduit. In some embodiments, at most 75% of the elements of the static mixer may be inserted into the Y-shaped conduit. In some embodiments, at most 50% of the elements of the static mixer may be inserted into the Y-shaped conduit. In some embodiments, at most 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 of the elements of the static mixer may be inserted into the Y-shaped conduit. [00116] In some embodiments, 2 of the 8 or more elements in a static mixer may be inserted into a Y-shaped conduit prior to the static mixer. In some embodiments, 6 of the 8 or more elements in a static mixer may be downstream from an outlet in the Y-shaped conduit. In some embodiments, 4 of the 12 or more elements in a static mixer may be inserted into a Y-shaped conduit prior to the static mixer. In some embodiments, 8 of the 12 or more elements in a static mixer may be downstream from an outlet in the Y-shaped conduit. In some embodiments, 2 of the 16 or more elements in a static mixer may be inserted into a Y-shaped conduit prior to the static mixer. In some embodiments, 4 of the 16 or more elements in a static mixer may be inserted into a Y-shaped conduit prior to the static mixer. In some embodiments, 12 of the 16 or more elements in a static mixer may be downstream from an outlet in the Y-shaped conduit. In Atty Dkt No.: 58530-742601 some embodiments, 14 of the 16 or more elements in a static mixer may be downstream from an outlet in the Y-shaped conduit. [00117] In some embodiments, any suitable number of elements in a static mixer may be inserted into a L-shaped conduit, based on for example, the length of the elements and the length of the L-shaped conduit. In some embodiments, at least 1 of the elements of the static mixer is inserted into the L-shaped conduit. In some embodiments, at least 2, 3, 4, 5, 6, 7, or 8 of the elements of the static mixer may be inserted into the L-shaped conduit. In some embodiments, at most 75% of the elements of the static mixer may be inserted into the L-shaped conduit. In some embodiments, at most 50% of the elements of the static mixer may be inserted into the L-shaped conduit. In some embodiments, at most 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 of the elements of the static mixer may be inserted into the L-shaped conduit. [00118] In some embodiments, 2 of the 8 or more elements in a static mixer may be inserted into a L-shaped conduit prior to the static mixer. In some embodiments, 6 of the 8 or more elements in a static mixer may be downstream from an outlet in the L-shaped conduit. In some embodiments, 4 of the 12 or more elements in a static mixer may be inserted into a L-shaped conduit prior to the static mixer. In some embodiments, 8 of the 12 or more elements in a static mixer may be downstream from an outlet in the L-shaped conduit. In some embodiments, 2 of the 16 or more elements in a static mixer may be inserted into a L-shaped conduit prior to the static mixer. In some embodiments, 4 of the 16 or more elements in a static mixer may be inserted into a L-shaped conduit prior to the static mixer. In some embodiments, 12 of the 16 or more elements in a static mixer may be downstream from an outlet in the L-shaped conduit. In some embodiments, 14 of the 16 or more elements in a static mixer may be downstream from an outlet in the L-shaped conduit. [00119] In some embodiments, each element of the static mixer may have a length less than about 2 in, 1.5 in, 1.2 in, 1 in, 0.9 in, 0.8 in, 0.7 in, 0.6 in, 0.5 in, 0.4 in, 0.3 in, 0.2 in, 0.19 in, 0.18 in, 0.17 in, 0.16 in, 0.15 in, 0.14 in, 0.13 in, 0.12 in, 0.11 in, or 0.10 in. In some embodiments, each element of the static mixer comprises a length of less than 50 mm, 40 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 5 mm, or 2.5 mm. In some embodiments, each element of the static mixer comprises a length of at least 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 5 mm, or 6 mm. In some embodiments, each element of the static mixer comprises a length of from about 1 mm to about 5 mm. In some embodiments, each element of the static mixer comprises a length of from about 1 mm to about 10 mm. In some embodiments, each element of the static mixer comprises a length of about 2.5 mm, as shown in FIGs.18A-C. In some Atty Dkt No.: 58530-742601 embodiments, each element of the static mixer comprises a length of about 3.8 mm, as shown in FIGs.19A-C. In some embodiments, each element of the static mixer comprises a length of about 3.2 mm, as shown in FIGs.21A-C. In some embodiments, each element of the static mixer comprises a length of about 1.5 mm, as shown in FIGs.22A-C. In some embodiments, each element of the static mixer comprises a length of about 4.8 mm, as shown in FIGs.23A-C. [00120] In some embodiments, each element of the static mixer may have a 45° twist. In some embodiments, each element of the static mixer may have a 90° twist, as shown in FIGs.22A- 22C. In some embodiments, each element of the static mixer may have a 135° twist. In some embodiments, each element of the static mixer may have a 180° twist, as shown in FIGs.9A-9C, 25 and 28. In some embodiments, each element of the static mixer may have a 225° twist. In some embodiments, each element of the static mixer may have a 270° twist, as shown in FIGs. 23A-23C, 26, and 27. In some embodiments, each element of the static mixer may have a 315° twist. In some embodiments, each element of the static mixer may have a -45° twist. In some embodiments, each element of the static mixer may have a -90° twist. In some embodiments, each element of the static mixer may have a -135° twist. In some embodiments, each element of the static mixer may have a -180° twist. In some embodiments, each element of the static mixer may have a -225° twist. In some embodiments, each element of the static mixer may have a - 270° twist. In some embodiments, each element of the static mixer may have a -315° twist. [00121] In some embodiments, each element of the static mixer makes a 270° twist every 1/8 inch. [00122] In some embodiments, the static mixer may have a 90° turn mixer, as shown in FIGs. 22A-22C. In some embodiments, the static mixer may have a 270° turn mixer, as shown in FIGs.23A-23C. [00123] In some embodiments, each element of the static mixer may have a cross section thickness less than about 5 mm, 4 mm, 3 mm, 2 mm, 1 mm, 0.5 mm, 0.4 mm, 0.3 mm, 0.2 mm, or 0.1 mm. In some embodiments, each element of the static mixer comprises a cross section thickness of at least 0.1 mm, 0.2 mm, 0.3 mm, 0.5 mm, 0.75 mm, 1 mm, 1.1 mm, 1.2 mm, 1.3 mm, 1.4 mm, or 1.5 mm. In some embodiments, each element of the static mixer comprises a cross section thickness of from about 0.1 mm to about 2 mm. In some embodiments, each element of the static mixer comprises a cross section thickness of from about 1 mm to about 2 mm. In some embodiments, each element of the static mixer comprises a cross section thickness of about 1 mm. In other embodiments, each element of the static mixer comprises a cross section thickness of about 1 mm. Atty Dkt No.: 58530-742601 [00124] In some embodiments, the static mixer may have one or more channel mixers. In some embodiments, the static mixer may have a 3-channel mixer. In some embodiments, the 3- channel mixer may be Y-shaped, as shown in FIGs.20A-20C. In some embodiments, the static mixer may have a 4-channel mixer. In some embodiments, the 4-channel mixer may be X- shaped, as shown in FIGs.21A-21C. [00125] In some embodiments, the systems herein provide for the requirement of reduced pressures as opposed to alternative systems, such as cross mixing systems. Reduced pressures may be useful to decrease the imposition of shear stress on the lipid nanoparticles and result in lower backpressure on pumping systems. Reduced pressures may be useful to allow adoption into larger scale production. In some embodiments, the pressure may be backpressure. In some embodiments, the systems provided herein reach a pressure of no more than 8 pounds per square inch gauge (psig), whereas cross mixing systems may reach a pressure of over 20 psig (e.g., 23.3 psig as described in the Examples). In some embodiments, the pressure of a system provided reaches no more than 20 psig (e.g., no more than 18 psig, 17 psig, 16 psig, 15 psig, 14 psig, 13 psig, 12 psig, 11 psig, 10 psig, 9 psig, or 8 psig). In some embodiments, the pressure of the system reaches at least 3 psig (e.g., at least 4 psig, 5 psig, 8 psig, 10 psig, 12 psig, 14 psig, or 16 psig). In some embodiments, the pressure of the systems provided herein reach from about 3 psig to about 20 psig. In some embodiments, the pressure of the systems provided herein reach from about 5 psig to about 15 psig, about 5 psig to about 10 psig, or about 6 psig to about 9 psig. Channel [00126] The present disclosure provides systems comprising a channel housing the static mixer. In some embodiments, the channel may be downstream of the conduit. In some embodiments, the channel may be upstream of the conduit. In some embodiments, the channel may be parallel to the conduit. [00127] The system may comprise a channel. The system may comprise a channel, wherein the channel houses a static mixer. The system may comprise a channel, wherein the channel houses an element of a static mixer. The system may comprise a channel, wherein the channel houses a plurality of elements of a static mixer. The system may comprise a channel, wherein the channel houses an element. The system may comprise a channel, wherein the channel houses a plurality of elements, In some embodiments, the channel provides a counterflow. The system may comprise a channel, wherein the nucleic acid and one or more lipid components are mixed to generate a mixture fluid. In some embodiments, the nucleic acid and one or more lipid components are mixed to generate a mixture fluid within the channel. Atty Dkt No.: 58530-742601 [00128] In some embodiments, the channel may have a length less than about 1000 millimeter (mm), 500 mm, 300 mm, 200 mm, 100 mm, 90 mm, 80 mm, 70 mm, 60 mm, 50 mm, 45 mm, 40 mm, 35 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 9 mm, 8 mm, 7 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, or 1 mm. In some embodiments, the channel comprises a length of at least 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 15 mm, 20 mm, 25 mm, or 30 mm. In some embodiments, the channel comprises a length of from about 5 mm to about 80 mm. In some embodiments, the channel comprises a length of from about 5 mm to about 40 mm. In some embodiments, the channel comprises a length of from about 10 mm to about 35 mm. In some embodiments, the channel may have a length less than about 10 inches (in), 5 in, 4 in, 3 in, 2 in, 1.5 in, 1.2 in, 1 in, 0.9 in, 0.8 in, 0.7 in, 0.6 in, 0.5 in, 0.4 in, 0.3 in, 0.2 in, or 0.1 in. In some embodiments, the channel has a length of about 4.8 mm. [00129] In some embodiments, the channel may have an inner diameter of less than about 5 in, 4 in, 3 in, 2 in, 1 in, ¾ in, ½ in, 3/8 in, ¼ in, 3/16 in, 1/8 in, 1/16 in, or 1/32 in. In some embodiments, the channel comprises an inner diameter of less than about 127 mm, 102 mm, 76 mm, 50 mm, 25 mm, 19 mm, 13 mm, 9.5 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, 1.5 mm, 1 mm, or 0.75 mm. In some embodiments, the channel comprises an inner diameter of at least 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 4 mm, or 5 mm. In some embodiments, the channel comprises an inner diameter of from about 0.5 mm to about 20 mm. In some embodiments, the channel comprises an inner diameter of from about 1 mm to about 10 mm. In some embodiments, the channel comprises an inner diameter of from about 1 mm to about 5 mm. In some embodiments, the channel comprises an inner diameter of about 3 mm. [00130] In some embodiments, the channel may be cylindrical, as shown in FIG. 25. In some embodiments, the channel may be square shaped, as shown in FIGs.26-28. In some embodiments, the channel may be rectangular shaped. In some embodiments, the channel may be any shaped. [00131] In some embodiments, the channel may have a twist at a length every less than about 2 in, 1.5 in, 1.2 in, 1 in, 0.9 in, 0.8 in, 0.7 in, 0.6 in, 0.5 in, 0.4 in, 0.3 in, 0.2 in, 0.19 in, 0.18 in, 0.17 in, 0.16 in, 0.15 in, 0.14 in, 0.13 in, 0.12 in, 0.11 in, or 0.10 in. In some embodiments, the channel may have a twist at a length every less than 50 mm, 40 mm, 30 mm, 25 mm, 20 mm, 15 mm, 10 mm, 5 mm, or 2.5 mm. In some embodiments, the channel may have a twist at a length every at least 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 5 mm, or 6 mm. In some embodiments, the channel may have a twist at a length every about 1 mm to about 5 mm. In Atty Dkt No.: 58530-742601 some embodiments, the channel may have a twist at a length every about 1 mm to about 10 mm. In some embodiments, the channel may have a twist at a length every 1/8 in. [00132] In some embodiments, the channel may have a plurality of twists. In some embodiments, the channel may have a 45° twist. In some embodiments, the channel may have a 90° twist. In some embodiments, the channel may have a 135° twist. In some embodiments, the channel may have a 180° twist. In some embodiments, the channel may have a 225° twist. In some embodiments, the channel may have a 270° twist. In some embodiments, the channel may have a 315° twist. In some embodiments, the channel may have a -45° twist. In some embodiments, the channel may have a -90° twist. In some embodiments, the channel may have a -135° twist. In some embodiments, the channel may have a -180° twist, as shown in FIG.26D, 27D, and 28D. In some embodiments, the channel may have a -225° twist. In some embodiments, the channel may have a -270° twist. In some embodiments, the channel may have a -315° twist. [00133] In some embodiments, the channel is square shaped and makes a -180° twist every 1/8 inch. Pump [00134] The present disclosure provides systems comprising a pump. The pump may be configured to provide one or more components of the lipid nanoparticle at a specified flow rate, such as a flow rate described elsewhere herein. In some embodiments, the pump is configured to provide the nucleic acid at a specified flow rate. In some embodiments, the pump is configured to provide the one or more lipids at a specified flow rate. In some embodiments, the pump is configured to provide the cationic ionizable lipid at a specified flow rate. In some embodiments, the pump is configured to provide the phospholipid at a specified flow rate. In some embodiments, the pump is configured to provide the polymer conjugated lipid at a specified flow rate. In some embodiments, the pump may be a peristaltic pump. In some embodiments, the pump may be a high performance liquid chromatography (HPLC) pump. In some embodiments, the pump may be a Levitronix pump. In some embodiments, the pump may be a Knauer pump. In some embodiments, the pump may be a Knauer BlueShadow pump. Generating Lipid Nanoparticles [00135] The present disclosure provides systems and methods for generating lipid nanoparticles. Provided herein are methods of using any of the systems provided herein, such as to prepare (generate) lipid nanoparticles. Atty Dkt No.: 58530-742601 [00136] In some embodiments, the methods provided herein comprise providing a system, such as a system provided elsewhere herein. In some embodiments, as described elsewhere herein, the system comprises a conduit. In some instances, the conduit comprises a first inlet. In some embodiments, the conduit comprises a second inlet. In some embodiments, the conduit comprises a third inlet. In some embodiments, the conduit comprises a fourth inlet. In some embodiments, the conduit comprises an outlet. In some embodiments, the system comprises a static mixer. In some embodiments, the conduit comprises a channel. [00137] In some embodiments, the methods provided herein comprise introducing a nucleic acid to the first inlet. In some embodiments, the methods provided herein comprise introducing one or more lipid components of the lipid nanoparticle to the second inlet. The systems or methods may comprise flowing a (e.g., first) solution comprising nucleic acid into a first inlet of a conduit, and a (e.g., second) solution comprising one or more lipid components into a second inlet of the conduit. In some embodiments, the first solution comprises any nucleic acid described elsewhere herein. In some embodiments, the second solution comprises one or more lipid components as described elsewhere herein. [00138] In some embodiments, the methods provided herein comprise introducing a nucleic acid to the first inlet or the second inlet or the third inlet or the fourth inlet. In some embodiments, the methods provided herein comprise introducing one or more lipid components of the lipid nanoparticle to the first inlet or the second inlet or the third inlet or the fourth inlet. The systems or methods may comprise flowing a (e.g., first) solution comprising nucleic acid into a first inlet or a second inlet or a third inlet or a fourth inlet of a conduit, and a (e.g., second) solution comprising one or more lipid components into the first inlet or the second inlet or the third inlet or the fourth inlet of the conduit. In some embodiments, the first solution comprises any nucleic acid described elsewhere herein. In some embodiments, the second solution comprises one or more lipid components as described elsewhere herein. [00139] In some embodiments, the methods provided herein comprise joining the nucleic acid and the one or more lipid components at the outlet. The first inlet and the second inlet may join into an outlet, such as an outlet described elsewhere herein, wherein the nucleic acid and the one or more lipid components are joined in the outlet. [00140] In some embodiments, the systems or methods may further comprise a static mixer. In some embodiments, the nucleic acid and the one or more lipid components are mixed within the static mixer to generate lipid nanoparticles. In some embodiments, the systems or methods may further comprise a static mixer, such as a static mixer described elsewhere herein, wherein Atty Dkt No.: 58530-742601 the nucleic acid and the one or more lipid components are mixed in a mixture fluid to generate lipid nanoparticles. [00141] In some embodiments, the systems or methods may further comprise a channel. In some embodiments, the nucleic acid and the one or more lipid components are mixed within the channel to generate lipid nanoparticles. In some embodiments, the systems or methods may further comprise a channel, such as a channel described elsewhere herein, wherein the nucleic acid and the one or more lipid components are mixed in a mixture fluid to generate lipid nanoparticles [00142] The systems and methods may generate lipid nanoparticles with a low polydispersity index, wherein the polydispersity index may be no greater than about 0.2. The systems and methods may generate lipid nanoparticles no greater than about 90 nanometers (nm). The systems and methods may generate mixture fluids with a Reynolds number no greater than about 1000. The systems and methods may have a ratio between a flow rate of the lipid component in the first inlet and a flow rate of the nucleic acid in the second inlet about 1:2. The systems and methods may have an encapsulation efficiency no less than about 90%. [00143] The conduit may comprise a first inlet, a second inlet, an outlet. The conduit may further comprise a junction site downstream of the first inlet and the second inlet. The method may comprise providing a static mixer. The method may comprise providing a static mixer, wherein the static mixer is disposed at a distal end of the junction site. The method may further comprise introducing a nucleic acid to the first inlet. The method may further comprise introducing one or more lipid components of the lipid nanoparticle to the second inlet. The method may further comprise joining the nucleic acid and the one or more lipid components at the outlet. The method may further comprise using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid. The method may further comprise using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid. Wherein the nucleic acid and the one or more lipid components may not contact each other prior to entering the junction site. [00144] The conduit may comprise a first inlet, a second inlet, a third inlet, a fourth inlet, an outlet. The method may comprise providing a static mixer. The method may further comprise introducing a nucleic acid to the first inlet or the second inlet or the third inlet or the fourth inlet. The method may further comprise introducing one or more lipid components of the lipid nanoparticle to the first inlet or the second inlet or the third inlet or the fourth inlet. The method may further comprise joining the nucleic acid and the one or more lipid components at the outlet. Atty Dkt No.: 58530-742601 The method may comprise providing a static mixer, wherein the static mixer may be configured to mix the nucleic acid and the one or more lipid components, wherein the static mixer comprises a plurality of elements, wherein each element of the static mixer makes a 270° twist every 1/8 inch. The method may further comprise providing a channel housing the static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the channel is square shaped and makes a -180° twist every 1/8 inch. [00145] In some embodiments, the mixture fluid may have a polydispersity index (PDI) no greater than about 0.5, 0.4, 0.3, 0.25, 0.20, 0.19, 0.18, 0.17, 0.160.15, 0.14, 0.13, 0.12, 0.11, 0.10, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or 0,01. In some embodiments, the mixture fluid comprises a polydispersity index (PDI) of at least 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09.0.10, 0.11, 0.12, 0.13, 0.14, or 0.15. In some embodiments, the PDI may be about 0.05 to about 0.5. In some embodiments, the PDI may be about 0.05 to about 0.25. In some embodiments, the PDI may be about 0.05 to about 0.2. In some embodiments, the PDI may be about 0.1 to about 0.5. In some embodiments, the PDI may be about 0.1 to about 0.25. In some embodiments, the PDI may be about 0.1 to about 0.2. In some embodiments, the PDI may be about 0.05 to about 0.16. In some embodiments, the PDI may be about 0.09 to about 0.12. The PDI may be measured using light scattering techniques, such as dynamic light scattering (DLS), or by electron microscopy (e.g., scanning electron microscopy or transmission electron microscopy). [00146] In some embodiments, the mixture fluid may have a Reynolds number of less than about 2500, 1500, 1200, 1000, 900, 800, 700, 600, 500, 400, 300, 200, or 100. In some embodiments, the mixture fluid may have a Reynolds number of at least 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000. In some embodiments, the Reynolds number is from about 100 to about 1200. In some embodiments, the Reynolds number is from about 100 to about 300. In some embodiments, the Reynolds number is from about 600 to about 900. In some embodiments, the Reynolds number is from about 600 to about 1000. In some embodiments, the Reynolds number is from about 600 to about 700. [00147] In some embodiments, the Reynolds number is calculated using the formula Re = (ρuL)/µ, wherein ρ is the density of the fluid, u is the flow speed, L is the characteristic linear dimension, and µ is the dynamic viscosity of the fluid. [00148] In some embodiments, increasing ρ, the density of the fluid, increases the Reynolds number. In some embodiments, decreasing ρ, the density of the fluid, decreases the Reynolds number. In some embodiments, increasing ρ, the density of the fluid, decreases the size of the Atty Dkt No.: 58530-742601 resulting lipid nanoparticles. In some embodiments, decreasing ρ, the density of the fluid, increases the size of the resulting lipid nanoparticles. In some embodiments, an increase in a pressure difference increases ρ, the density of the fluid. In some embodiments, an increase in a pressure difference increases the Reynolds number. In some embodiments, a decrease in a pressure difference decreases ρ, the density of the fluid. In some embodiments, an increase in a pressure difference decreases the size of the resulting lipid nanoparticles. In some embodiments, a decrease in a pressure difference increases the size of the resulting lipid nanoparticles. [00149] In some embodiments, increasing u, the flow speed, increases the Reynolds number. In some embodiments, decreasing u, the flow speed, decreases the Reynolds number. In some embodiments, modifying u, the flow speed, controls the size of the resulting lipid nanoparticle. In some embodiments, increasing u, the flow speed, decreases the size of the resulting lipid nanoparticles. In some embodiments, decreasing u, the flow speed, increases the size of the resulting lipid nanoparticles. In some embodiments, an increase in a pressure difference increases u, the flow speed. In some embodiments, an increase in a pressure difference increases the Reynolds number. In some embodiments, a decrease in a pressure difference decreases u, the flow speed. In some embodiments, a decrease in a pressure difference decreases the Reynolds number. In some embodiments, an increase in a pressure difference decreases the size of the resulting lipid nanoparticles. In some embodiments, a decrease in a pressure difference increases the size of the resulting lipid nanoparticles. [00150] In some embodiments, increasing L, the characteristic linear dimension, increases the Reynolds number. In some embodiments, decreasing L, the characteristic linear dimension, decreases the Reynolds number. In some embodiments, increasing L, the characteristic linear dimension, decreases the size of the resulting lipid nanoparticles. In some embodiments, decreasing L, the characteristic linear dimension, increases the size of the resulting lipid nanoparticles. In some embodiments, changing L, the characteristic linear dimension, and changing u, the flow speed, maintain the Reynolds number. In some embodiments, L, the characteristic linear dimension is a channel size. [00151] In some embodiments, increasing µ, the dynamic viscosity of the fluid, decreases the Reynolds number. In some embodiments, decreasing µ, the dynamic viscosity of the fluid, increases the Reynolds number. In some embodiments, increasing µ, the dynamic viscosity of the fluid, increases the size of the resulting lipid nanoparticle. In some embodiments, decreasing µ, the dynamic viscosity of the fluid, decreases the size of the resulting lipid nanoparticle. Atty Dkt No.: 58530-742601 [00152] In some embodiments, increasing the Reynolds number decreases the particle size of the lipid nanoparticle. In some embodiments, decreasing the Reynolds number increases the particle size of the lipid nanoparticle. In some embodiments, increasing the Reynolds number increases the particle size of the lipid nanoparticle. In some embodiments, decreasing the Reynolds number decreases the particle size of the lipid nanoparticle. In some embodiments, the Reynolds number remains the same as the particle size of the lipid nanoparticle increases. In some embodiments, the Reynolds number remains the same as the particle size of the lipid nanoparticle decreases. [00153] In some embodiments, as the Reynolds number increases, the particle size (e.g., diameter) of the lipid nanoparticles decreases, such as shown in FIG.3A. In some embodiments, as the Reynolds number increases the PDI of the lipid nanoparticles remains about the same, such as shown in FIG.3B. In some embodiments, provided herein are methods of generating lipid nanoparticles, wherein the particle size is selected by increasing or decreasing the Reynolds number. In some embodiments, the method comprises increasing the Reynolds number to decrease particle size. In some embodiments, the method comprises decreasing the Reynolds number to increase particle size. In some embodiments, the relationship between the Reynolds number and particle size is an inverse relationship. [00154] In some embodiments, the lipid nanoparticle may have a size no greater than 500 nanometers (nm), 400 nm, 300 nm, 200 nm, 100 nm, 90 nm, 80 nm, 70 nm, 60 nm, 50 nm, 40 nm, 30 nm, 20 nm, 10 nm, 5 nm, or 1 nm. In some embodiments, the lipid nanoparticle comprise a size of at least 10 nm, 20 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 85 nm, 90 nm, 100 nm, 105 nm, 110 nm, 120 nm, 140 nm, 160 nm, 180 nm, or 200 nm. In some embodiments, the lipid nanoparticle comprises a size of from about 50 nm to about 300 nm. In some embodiments, the lipid nanoparticle comprises a size of from about 50 nm to about 100 nm. In some embodiments, the lipid nanoparticle comprises a size of from about 100 nm to about 200 nm. In some embodiments, the lipid nanoparticle size is measured by light scattering techniques (e.g., dynamic light scattering), or by electron microscopy (e.g., scanning electron microscopy or transmission electron microscopy). [00155] In some embodiments, such as shown in FIG. 6, particle size (e.g., diameter) remains the same after mixing in the static mixer and after dilution. In some embodiments, dilution comprises dilution with an aqueous solvent. In other embodiments, dilution comprises dilution with ethanol (e.g., such as to 12.5 v/v%). Atty Dkt No.: 58530-742601 [00156] In some embodiments, the methods provided herein comprise introducing ethanol after the (e.g., first and second) solutions enter and leave the static mixer (e.g., post-mixer). In some embodiments, the amount particle size and polydispersity index (PDI) are influenced by the amount of ethanol introduced post mixer. In some embodiments, the amount of ethanol introduced post-mixer is at least 10%. In some embodiments, the amount of ethanol introduced post-mixer is at least about 12%, 15%, 17%, 25%, 33%, 40%, 45%, 50%, or at least 55%. In some embodiments, the characteristics of the resulting particles with varying amounts of ethanol introduced post-mixer are shown in Table 4. In some embodiments, the polydispersity is <0.15 for lipid nanoparticles diluted with at least 25% ethanol. In some embodiments, the polydispersity is about 0.2 or higher for lipid nanoparticles diluted with l7% or less ethanol post- mixer. In some embodiments, the relationship between particle size and amount of ethanol is parabolic, with the lowest particle sizes exhibited after introduction of 25% ethanol post-mixer, such as described in Table 4. [00157] In some embodiments, an encapsulation efficiency may be no less than about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, an encapsulation efficiency may be no more than about 99.9%, 99.5%, 99%, 98%, 96%, 94%, 90%, or 85%. In some embodiments, the encapsulation efficiency may be from about 40% to about 99.9%, about 50% to about 99%, about 60% to about 99%, about 70% to about 99%, about 80% to about 99%. In some embodiments the encapsulation efficiency is from about 90% to about 99%, such as shown in FIG.7C. [00158] In some embodiments, the lipid nanoparticles generated by the methods provided herein using the systems provided herein are stable for at least 24 hours (e.g., at least 48 hours, 1 week, 1 month, 3 months, or at least 6 months). In some embodiments, the lipid nanoparticles are stable for from about 24 hours to about 6 months, from about 24 hours to about 3 months, about 1 week to about 6 months, or from about 1 month to about 6 months. In some embodiments, the lipid nanoparticles are stable at a temperature of from about 2°C to about 8°C. In some embodiments, the lipid nanoparticles are stable at a temperature of about -80°C. [00159] In some embodiments, a time of mixing may be no less than about 5 milliseconds (ms), 10 ms, 15 ms, 20 ms, 25 ms, 30 ms, 35 ms, 40 ms, 45 ms, 50 ms, 55 ms, or 60 ms. In some embodiments, a mixing time may be no more than about 5 ms, 10 ms, 15 ms, 20 ms, 25 ms, 30 ms, 35 ms, 40 ms, 45 ms, 50 ms, 55 ms, or 60 ms. In some embodiments, the mixing time may be from about 5 ms to about 40 ms, such as shown in FIGs.17A-B. Atty Dkt No.: 58530-742601 [00160] While various embodiments of the disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the disclosure. It should be understood that various alternatives to the embodiments of the disclosure described herein may be employed. [00161] As used herein and in the appended claims, the singular forms “a,” “and,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “an agent” includes a plurality of such agents, and reference to “the cell” includes reference to one or more cells (or to a plurality of cells) and equivalents thereof known to those skilled in the art, and so forth. When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formulae, all combinations and subcombinations of ranges and specific embodiments therein are intended to be included. The term “about” when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range may vary between 1% and 15% of the stated number or numerical range. The term “comprising” (and related terms such as “comprise” or “comprises” or “having” or “including”) is not intended to exclude that in other certain embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, described herein, may “consist of” or “consist essentially of” the described features. [00162] Whenever the term “at least,” “greater than,” or “greater than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “at least,” “greater than” or “greater than or equal to” applies to each of the numerical values in that series of numerical values. For example, greater than or equal to 1, 2, or 3 is equivalent to greater than or equal to 1, greater than or equal to 2, or greater than or equal to 3. [00163] Whenever the term “no more than,” “less than,” or “less than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “no more than,” “less than,” or “less than or equal to” applies to each of the numerical values in that series of numerical values. For example, less than or equal to 3, 2, or 1 is equivalent to less than or equal to 3, less than or equal to 2, or less than or equal to 1. Atty Dkt No.: 58530-742601 Examples Example 1: Static Mixer for Lipid Nanoparticle Production [00164] Cross-tee mixing is a mixing method that requires high energy input under turbulent flow conditions. Furthermore, when operating under high flow conditions, high back-pressure is exhibited, necessitating the use of specific pumps to manage the increased pressure. Alternative methods utilizing lower flow and back-pressure may be desirable to gain better control over the physicochemical properties of the drug product. Transitioning to lower flow and lower back- pressure regime could provide a broader selection of pump options and a better control on lipid nanoparticle (LNP) formation. Static mixers may prove to be a suitable choice for mixing processes. Methods [00165] Lipid components for the desired formulation were dissolved in ethanol to achieve the appropriate concentration. For further details on lipid formulations, see Tables 8 and 11. For “empty” LNP tests, the aqueous mRNA phase consisted of 15 mM citrate pH 4 buffer with no mRNA added. For tests using mRNA, they were formulated according to Table 9. Static mixers of varying length, diameter, and architecture were purchased from Cole Palmer, and are depicted in FIGs.1A-I. Each system was run with a 2:1, 3:1, or 5:1 buffer:lipid flow rate ratio (FRR) at varying total flow rate (TFR) corresponding to Reynolds numbers at static mixer outlet. These total flow rates are shown in Tables 1, 3, 5, and 6. Samples made with 2:1 flow rate ratios also had an inline buffer dilution at the same flow rate as the lipid flow rate. Some systems were also tested with food coloring dissolved in ethanol and water to simulate and visualize the mixing properties of each system. All Reynolds numbers were calculated using the diameter of the static mixer outlet. [00166] Mixing was achieved using three Chemyx Fusion 6000x syringe pumps and syringes ranging in size from 20 to 100 mL. For mRNA-encapsulating formulations that did proceed to buffer exchange after water dilution, buffer exchange was performed using Cytiva PD-10 columns. Each column was equilibrated 4 times with 4 mL of storage buffer for that formulation. 2.5 mL of formulation was loaded onto each column and eluted with 3.5 mL of storage buffer. The formulations were concentrated to a target mRNA concentration of 1 mg/mL via ultracentrifugation using Amicon Ultra-15 centrifugal filters and an optional buffer dilution to the target formulation volume at 1 mg/mL. Formulations were then filtered with a single PES syringe filter. Atty Dkt No.: 58530-742601 [00167] Particle size distribution data was obtained via dynamic light scattering (DLS). Formulation samples were diluted 33-fold to a nominal concentration of 0.03 mg/mL in the diluent of 0.2X phosphate-buffered saline (PBS). In-process formulation samples were run on the Wyatt DynaPro Plate Reader III. Particle size distribution data after overnight sample storage was measured on both the Wyatt plate reader and the Malvern Zetasizer Ultra-Red. Particle size (nm) and polydispersity index (PDI) values from the Wyatt plate reader are reported as the mean values of triplicate measurements, whereas there were no replicate measurements run on the Malvern Zetasizer. Results and Discussion Mixer Schematics [00168] FIGs. 1A-I show schematics of each mixing system initially tested, with blue representing the organic lipid inlet and yellow representing the aqueous mRNA inlet. Systems A and B (FIGs.1A-B) both use the same mixing architecture, a Koflo 24-element 1/8” ID disposable static mixer with a polypropylene tube. System C (FIG.1C) uses a Koflo 12-element ¼” OD mixing element fitted into ¼” ID platinum cured silicone tubing. Systems D and E both are manipulated versions of system B, with system D (FIG.1D) having the mixing architecture pushed partway into the Y, and with system E (FIG.1E) having a makeshift flow splitter incorporated by way of a ziptie inserted into the Y. Both of these customizations were made in attempts to minimize the mixing occurring in the Y, in order to better test the actual mixing architecture’s ability to form nanoparticles. System F is similar to System B, but with only 12 mixing elements. System G (FIG.1G) is a custom design with three 12-element 1/8” mixers fit next to each other in a 3/16” silicone tube. Systems H and I (FIGs.1H-I) are both custom systems with 1/8” mixers pushed into the flow joints such that the mixer starts upstream of the point at which the fluids meet, with H having a tee joint and I having a Y joint. Visualizing Mixing with Food Dye [00169] FIG. 2 shows images of systems A, B, and C while being tested with food dye feedstocks. The blue food dye is dissolved in ethanol, while the yellow food dye is dissolved in MilliQ water, to best illustrate lipid-mRNA mixing. Images for systems A and B are shown at joint outlet Reynolds numbers of 25, 50, and 100, while system C is just shown at Re of 100. The mixing quality of system C may be seen to be significantly lower than in systems A and B, which both seem to achieve a consistent green color after a few mixing elements at all tested Re. It is worth noting that clearance of air from the mixing systems was difficult, with bubbles Atty Dkt No.: 58530-742601 consistently travelling through the mixer through the entire process. This was more of an issue with the smaller-diameter mixer. Day 1 System Runs [00170] On day 1 of testing, Systems A-E were tested with food dye, and after visually evaluating mixing performance, Systems B, D, and E were run under multiple conditions. While FRR was held constant at 1:2 lipid:buffer ratio, Reynolds number was changed by varying the total flow rate of the system. All results from day 1 of testing are shown in Table 1A and Table 1B for LNP 5, with generally high particle sizes compared to the control system. Out of the mixers tested on this day, System B at Re of 621 and systems D and E at Re of 276 performed favorably. Table 1A Table 1B [00171] For System B, increasing Reynolds number decreased the resultant LNP size. This trend is plotted in FIGs.3A-B. This trend follows the inverse relationship between flow rate and time of mixing, low values of the latter commonly being correlated with smaller nanoparticle Atty Dkt No.: 58530-742601 formation. Particle sizes 15 minutes post-mix and directly following dilution are shown in FIG. 3A, while the PDI for the readings in FIG.3A are shown in FIG.3B. Day 2 System Runs [00172] On day 2 of testing, new system designs were tested, as well as FRRs different than the standard 1:2 lipid:buffer ratio. All but the last three tests were conducted with leftover LNP 3 lipids (a five-lipid composition), in order to conserve materials. Table 2A and Table 2B shows all results from this day of testing, including two control runs conducted on the tee mixer (one at normal Re, another at Re comparable to most of the static mixer tests). System B was tested again with higher citrate inline ratios (1:5 instead of 1:2), which resulted in larger PS than the previous tests at 1:2 FRR. System B with 1:5 FRR and Re of 621 was also tested with immediate water dilution rather than 30-minute delayed water dilution, a test that showed PS and PDI independence to this change. [00173] New systems with iterated designs were also tested, including systems F, G, and H, all at 1:2 FRR and Re of 621. Between these three systems, F and H both performed similarly well and better than any previously performed static mixer test. System F did, however, result in a high PDI after dilution, while System H did not. System H was also tested at a FRR of 1:3 while keeping Re constant, showing no difference in PS and a minor increase in PDI. [00174] Finally, Systems F, H, and I were tested with LNP 5 due to the favorable results thus far (System I hadn’t been tested yet but was a variation of system H, which performed well). All systems used FRRs of 1:3, and System F performed worst both in terms of PS and PDI. Systems H and I performed relatively well compared to the other static mixing experiments, resulting in particle sizes of 115 nm and 102 nm post-dilution, respectively, as well as PDI at or below 0.15. It is worth noting that System I was also tested at a higher Re, due to its smaller Y bore size. This difference in Re could be the cause of the lower PS compared to System H, due to higher mixing efficiency. Table 2A Atty Dkt No.: 58530-742601 Table 2B Day 3 System Runs [00175] Given the results from Day 1 and Day 2, follow-up experiments were conducted to optimize the design of the most promising system, System H (FIG.1H). From this system, six new test systems were devised, one of which being a negative test, and will be referred to as Systems J-O. All architectures used LNP 5 lipid and buffer, and visual representations of these systems are shown in FIGs.4A-F. While the same mixing architecture is tested in Systems J-N (FIGs.4A-E), FRR, number of mixing elements, orientation of feed inlets, and residence length Atty Dkt No.: 58530-742601 were varied to observe their effects on post-mix and post-dilution particle size. Table 3A and Table 3B summarizes the experimental parameters and results from these tests. Table 3A Table 3B [00176] When directly comparing the newly tested systems, one can see that favorable performance was achieved in Systems J, L, and N (FIG.4A, C, E). All three of these systems utilize the helical mixer with approximately two mixing elements inserted into the mixing tee. These three systems all also have an EtOH concentration post-mix of 25%, though system L (FIG.4C) achieves this same percentage of EtOH after the initial mixing with no later in-line dilution. Systems J and N (FIG.4A, E) performed similarly even though the latter system has half as many mixing elements. This result may support the hypothesis that the most important emulsion formation occurs quickly at the onset of mixing. A higher flow rate ratio (System M (FIG.4D)) resulted in large particles as well as high PDI. Overall, the best designs of static mixing systems were able to mimic the tee mixer control with comparable PS and PDI at a Re of Atty Dkt No.: 58530-742601 1000, though this did require a much higher flow rate due to the relatively increased inner diameter of the tubing. [00177] Systems J and K (FIGs. 4A-B) tested the feedstock orientation, with System J having the lipid inlet at the junction and system K having the lipid inlet at the tee through line. System J showed better results than System K, with lower PS and comparable PDI. This result is possibly because System J has the higher flow-rate inlet in the through line, potentially causing less back pressure. FIGs.5A-B shows bar graphs of both the particle diameters post-dilution (FIG.5A) and PDI post-dilution (FIG.5B). Day 4 System Runs [00178] Following the tests on Day 3, a test of higher ethanol amounts at static mixer outlet (pre-inline dilution) was conducted in order to find the optimum post-mixer ethanol concentration (ethanol (v/v) percentage) for the static mixing architecture. System J (FIG.4A) was used for these tests, along with LNP 5 lipid and buffer, and a Reynolds number of approximately 1000 at mixing outlet was maintained across all tested conditions, assuming negligible change to Reynolds number by bulk viscosity and density (actual Re ranges from 920 to 1001). For citrate dilution flow rates unable to be achieved by the Chemyx syringe pumps, a peristaltic pump was used instead (Masterflex Model 77200-30). All relevant mixing parameters are shown in Table 4. All samples were diluted to 12.5 (v/v)% ethanol 30 minutes post-mix, and the citrate dilution flow rate was determined such that the post-inline dilution ethanol (v/v)% was maintained at 25%. Some samples from Table 3A and Table 3B are included in Table 4A and Table 4B for comparison. Table 4A Atty Dkt No.: 58530-742601 Table 4B [00179] Table 4B also shows particle size and PDI post-mix and post-dilution for all samples. PDI was generally low (<0.15) for formulations with at least 25% ethanol post-mixer, with 17% ethanol post-mixer resulting in PDI around 0.2. Particle size showed a parabolic relationship to ethanol amount post-mixer, with sizes ranging from ca.85 to 160 nm and a minimum particle size reached around 33% ethanol post-mixer. The 1:2 FRR being favorable aligns the static mixer with previous mixers, as this 1:2 ratio has previously been found to be a favorable mixing ratio for both tee and cross-tee mixers. FIG.6A shows the particle diameters plotted against ethanol post-mixer, and FIG.6B shows the PDI. Day 5 System Runs [00180] After analyzing results from the previous three sets of experiments, the design from System N (FIG.4E) was further analyzed, due to its achievement of low particle size and PDI with fewer required mixing elements. System N was then tested with an mRNA formulation instead of only lipid and buffer, in order to evaluate encapsulation rate and storage stability Atty Dkt No.: 58530-742601 parameters previously unexamined on this architecture. LNP 5-CFTR was used, and flow rate ratios of 1:2 and 1:3 were tested for the static mixing setup. A tee mixer control was also run with the same formulation and a 1:2 flow rate ratio, and all systems were run at Re of 1000 at mixing outlet. All experimental flow parameters are shown in Table 5. Table 5 [00181] Each formulation was made and processed through small-scale buffer exchange (PD 10 columns), then subsequent centrifuge-concentration (Amicon tubes), and filtration. Finally, storage stability was tested by storing aliquots at either 2-8°C or -80°C for over 24 hours. After each processing and storage step, particle size and size distribution were analyzed, the values being reported in Table 6A and Table 6B. Table 6A Table 6B Atty Dkt No.: 58530-742601 [00182] Particle sizes were consistently the lowest in the control system, with PS ranging from 61 to 65 nm, and PDI under 0.1. The static mixer test with a FRR of 1:2 generally performed better than the 1:3 FRR, with the former achieving PDI consistently at or below 0.1 and PDI ranging from 72 to 77 nm. The static mixer with 1:3 FRR interestingly had a particle size that steadily increased during processing, starting at 78 nm post-mix and increasing to 101 nm post-concentration. Post-concentration, the particle size increased by 14 nm. Not shown in Table 6A and Table 6B is the encapsulation efficiency, which was between 98 and 99% after both storage conditions for the control and 1:2 FRR static mixer conditions, and was much lower at 66-68% for the 1:3 FRR condition. It is also worth noting that in this study the 1:3 FRR condition had a different post-mix ethanol concentration (20% rather than 25%) due to it still having in-line citrate dilution. This was accounted for in the dilution step by diluting with less water to achieve the same 12.5% ethanol post-dilution as was achieved by 1:1 dilution in the control and 1:2 FRR static mixer systems. Day 6 System Runs [00183] After concluding in the previous tests that the System N static mixer architecture with 1:2 FRR performs well with LNP 5-CFTR (PDI < 0.1, PS between 60 and 85 nm, and encapsulation > 97%), a final control test of the system with and without the mixer elements was completed. A second mixing system was constructed identical to System N (same tubing lengths, attachments, and connectors) but without any included static mixing elements. This was called the “No Mixer” system, and was tested to verify that the mixing was occurring primarily due to the static mixing elements and not due to the junction of fluids (e.g., via jet impingement). The two systems were tested according to the flow parameters outlined in Table 7. Table 7 Atty Dkt No.: 58530-742601 [00184] Both systems were tested at the same flow parameters to identify mixing characteristics for systems at Re of 1000 with and without static mixing elements. The samples were processed in the same manner as the samples previously prepared (Day 5 of testing), and the results from processing are shown in Table 8A and Table 8B. Not shown in Table 8A or Table 8B is the encapsulation efficiency post-storage for each formulation. For the control system with no static mixing elements, an encapsulation efficiency of 49-51% was achieved, whereas with the 1:2 FRR static mixing system an encapsulation of 96% was achieved. It was noted that the 96% encapsulation efficiency was lower than usually seen with this static mixing system, and lower than is usually expected with an LNP 5-CFTR formulation. Table 8A Table 8B [00185] A general trend of lower performance can be seen in Table 8A and Table 8B for the control, no-mixer configuration. Particle sizes ranging from 186 to 235 nm were found during processing and post-storage, with maximum particle size seen after spin concentration and minimum particle size seen after refrigeration. PDI for the no-mixer system ranged between 0.09 and 0.22, values generally being elevated compared to most mixing systems with LNP 5-CFTR. The static mixer performed better in comparison to the no mixer system, with particle sizes ranging from 86 to 99 nm, with highest particle size seen post-mix and decreasing incrementally during processing. The static mixer sample showed the smallest particle size post-refrigeration. PDI was generally low for the static mixer sample, with values ranging from 0.06 to 0.14 and Atty Dkt No.: 58530-742601 peaking post-spin concentration. Once again, these values are not as ideal as those seen with the same formulation and conditions tested on previously (higher particle size and PDI, and lower encapsulation). However, this test does show that the mixing elements may directly increase the mixing performance of the system by drastically lowering particle size and increasing the encapsulation efficiency. Day 7 System Runs [00186] After the Day 6 control test showed that the static mixing elements provided a significant increase in mixing performance with LNP 5, other mRNA and lipid formulations were tested to evaluate how the static mixing architecture performs cross-platform. To do this, five distinct formulations with two different payloads were tested. For exact lipid compositions and mRNA batches for each formulation used in these tests, see Table 11. The experimental parameters for these tests are laid out in Table 9, which shows that three formulations using CFTR mRNA and two formulations using DNAI1 mRNA were made. All systems used the same mixing architecture (System N) and the same number of elements, as well as the same FRR, Reynolds number at bore outlet, and inlet flow rates. The main variants between the systems were lipid composition (see Appendix A), lipid:mRNA ratio, lipid and mRNA feedstock concentration, buffer strength, buffer pH, and final storage buffer composition. Table 9 Atty Dkt No.: 58530-742601 [00187] All size and distribution data are shown in Table 10A and Table 10B. LNP 1-CFTR resulted in the highest particle sizes and largest size distributions across the board, with sizes ranging from 84 to 106 nm and PDI ranging from 0.09 to 0.17, with maximum particle size post- dilution and maximum PDI post-mix. LNP 5-CFTR also resulted in large particles, but were smaller and less polydisperse than those of LNP 1-CFTR. Particle sizes for LNP 5 ranged from 82 to 93 nm, with PDI ranging from 0.08 to 0.17 (the latter an outlier post-spin concentration). Smallest particle sizes were seen post-storage. LNP 3-CFTR showed similar behavior in-process to LNP 5-CFTR, with particle size ranging from 82 to 95 nm, and PDI ranging from 0.08 to 0.13. Both DNAI1 formulations performed better than all three CFTR formulations, with LNP 2- DNAI1 having particle size range of 78-86 nm and PDI range of 0.02-0.15. Finally, LNP 4- DNAI1 had the lowest particle size range of the five formulations tested with a range of 63 to 73 nm and a PDI range of 0.06 to 0.16. [00188] With this cross-platform comparison it can be seen that the static mixer system can indeed make viable, though slightly larger, nanoparticles with low polydispersity (less than 0.2) and moderate particle size (62-87 nm post-storage). Not included in Table 10A and Table 10B are encapsulation efficiency values, which were generally above 97%, with a few exceptions. First, LNP 3-CFTR had a post-freeze/thaw encapsulation efficiency of 93%, which was peculiar since encapsulation efficiency post-refrigeration was 98.6%. Second, LNP 5-CFTR had an encapsulation efficiency of 94% post-refrigeration, which again was strange given its 97.8% post-freeze/thaw encapsulation efficiency. Lastly, LNP 2-DNAI1 had an encapsulation efficiency of ~90% after both storage methods. Atty Dkt No.: 58530-742601 Table 10A Table 10B [00189] To help draw conclusions from this cross-platform screening, it is important to compare these results to previous results using a mixer of established quality. To do this, the post-storage particle size, PDI, and encapsulation efficiency of the static mixing system were compared to previously formulated compositions made using a cross-tee mixer. It is worth noting that the compared LNP 4 formulation uses a different mRNA (H1N1 rather than DNAI1), and LNP 1 uses a tagged version of CFTR. The plotted comparisons are shown in FIGs.7A-C. One can see that particle diameters are generally higher with the static mixing architecture than when made with the cross-tee architecture. PDI is generally comparable between the two mixing systems, with static mixer having lower PDI in 5 of 9 of the comparisons. Encapsulation Atty Dkt No.: 58530-742601 efficiency is also comparable, with the largest deviations in LNP 1 and LNP 2, with the former having higher encapsulation efficiency when made with static mixer, and the latter having lower encapsulation efficiency when made with static mixer. Conclusion [00190] In this study different static mixing architectures were tested and iterated to work towards a new mixing geometry to replace a cross-tee mixer. This replacement should ideally be scalable and should decrease downstream particle aggregation seen with the current mixer. Across the board, initial static mixing systems exhibited higher particle size than control runs with tee mixer, even when the tee mixer was run at a comparable Reynolds number to the static mixers. This may be due to the mixing starting prior to the lipid and mRNA stocks encountering the static mixer architecture, a theory that resulted in the designs for Systems D, E, H, and I. Some of these systems did indeed result in lower PS than the first round of designs, notably Systems H and I, each of which having the mixer stretch into the lipid inlet prior to joining with the buffer stream. This optimization lowered the particle size from ca.200 to ca.100 nm, a 50% reduction. [00191] These systems were then iterated to give Systems J-O (FIGs. 4A-F), which gave much better results when tested at a Re of 1000, when compared to the previously tested static mixers. In these tests it was found that favorable particle diameter and PDI was found at 1:2 FRR with the higher-flow-rate aqueous inlet passing through the straight portion of the joint and the organic lipid inlet joining at the junction. Negligible differences were found between 8- and 16- element mixing systems, underlining the importance of initial mixing events on quality emulsion formation. All in all, the third round of static mixing tests showed great promise, with the single caveat of them needing to be run at slightly higher Re/flow rate in order to mix well. [00192] Following Day 3 of mixer testing, ethanol concentration post-junction was probed, and a favorable mixing ratio of 1:2 was found. It was then decided that further testing would be performed on System N due to its high performance with fewer elements. LNP 5-CFTR was tested using System N to compare to a control (tee mixer), which showed that the static mixing system performed favorably at a 1:2 lipid:mRNA flow rate ratio (FRR), while still making slightly larger particles than the tee mixer. On Day 5 of testing a true control system was manufactured and tested – a system with the same attachments, tubing lengths, and connectors as System N, but with no static mixing elements – in order to decouple mixing effects from the static mixing elements and the joining of fluid paths. From these tests it was found that the system may indeed require the static mixing elements to properly make particles smaller than Atty Dkt No.: 58530-742601 150 nm, and that without the mixing elements the mRNA encapsulation efficiency dropped below 60%. [00193] Finally, the static mixing system (System N) was characterized on five different mRNA platforms: LNP 1-CFTR, LNP 2-DNAI1, LNP 3-CFTR, LNP 4-DNAI1, and LNP 5- CFTR. When comparing the post-storage results from static mixing to those of analogous formulations made with the cross-tee mixing system, the static mixer generally produced particles with comparable encapsulation efficiency and PDI, but with particle diameters higher by an average of 14 nm. Table 11 Example 2: Static Mixer Mixing Simulation [00194] A mixing simulation using the Reynolds stress model between ethanol and water demonstrated lower maximum shear stress and system pressure in the static mixer than in the cross mixer, as shown in FIGs.15A-C. The static mixer had a length of 1 inch, and an inner diameter of about 1/8 inches. The static mixer contained 8 elements, in which each element was about 1/8 inches in length, and each element makes an about 180° twist, as shown in FIG.15A. The cross-section thickness of each element was about 0.0394 inches (1 mm), as shown in FIG. 15A. The ethanol flow rate was about 75.4 mL/min and the water flow rate was about 150.8 mL/min, as shown in FIG.15B. FIGs.16A-B Shows the comparative system, a standard about 0.5 mm inner-diameter cross mixer fluid path with three inlets, two of which have water flow rates of about 40 mL/min and one of which has an ethanol flow rate of about 40 mL/min. The outlet has an inner diameter of about 1/16 in. Both systems were simulated in Ansys Fluent, and the results were used to extract shear stress, total system pressure, and mass fraction of ethanol. Cross sections were taken about every 0.1 mm perpendicular to direction of fluid travel, and on Atty Dkt No.: 58530-742601 these cross-sections area-weighted average shear stress and standard deviation of ethanol mass fraction were calculated. The average shear stress term was used to evaluate the amount of shear stress each mixing architecture imposed on the mixing fluids, and the standard deviation of ethanol mass fraction was used to evaluate the homogeneity of the fluid, with low standard deviation representing high degree of mixing/homogeneity. Time was calculated in milliseconds from the fluid flow rates from each system and the cross-sectional areas of each evaluated cross- section. A time point of zero was set at the initial point of water-ethanol flow intersection for each mixing system. The results from these simulations are shown in FIGs.17A-C. Comparing ethanol mass fraction standard deviation results from the two different mixers (FIG.17A), the cross mixer is shown to have a faster decrease in standard deviation, indicating a faster mixing process when compared to the simulated static mixing system. However, when comparing area- averaged shear stress (FIG.17B), the cross mixer is shown to have much higher shear stress during the mixing process, with a maximum reaching almost about 1000 Pa, which is over five times the maximum shear stress reached by the static mixing system. Furthermore, when comparing the maximum total system pressures of the two systems (FIG.17C), one can see that the cross mixing system reaches a pressure of about 23.3 psig, while the static mixing system only reaches a pressure of about 7.7 psig. These results indicate that although the cross mixing system approaches a homogeneous system faster than the proposed static mixing system, the latter is far improved in its decreased imposition of shear stress on the simulated material, as well as in its lower system pressure, which should directly indicate lower backpressure imposed on pumping systems. Example 3: Static Mixer Designs [00195] A series of mixers were designed and 3D printed using Digital Light Processing (DLP), a resin-based additive manufacturing technique with high resolution and greater accuracy. Loctite 3955, FST, Black was selected as the mixer material, known for its chemical resistance to ethanol. FST composite resins are fire, smoke, and toxicity resistant. [00196] A Koflo analogue design (Design 1, No. A1) was made, modeled after the Koflo static mixers. The interior elements are helical elements with 180° twists (FIG.25A). The exterior housing comprises 2 inlets with 1/8 inch barbs that are 120° apart (FIG.25B), which is wider than the typical Y connection of a 60° separation to allow space for tubing attachments, and zip ties. The channel is cylindrical shape which houses the elements (FIG.25C). Atty Dkt No.: 58530-742601 [00197] Another design (Design 2, No. D10) was made, which included helical elements with 270° twists (FIG.26A). The exterior housing comprises 2 inlets (FIG.26B) and the channel is square shaped (FIG.26C) and comprise -180° twists (FIG.26D). [00198] Another design (Design 3, No. D7) was made, which included helical elements with 270° twists (FIG.27A). The exterior housing comprises 4 inlets (FIG.27B) and the channel is square shaped (FIG.27C) and comprise -180° twists (FIG.27D). [00199] Another design (Design 5, No. D6) was made, which included helical elements with 180° twists (FIG.28A). The exterior housing comprises 4 inlets (FIG.28B) and the channel is square shaped (FIG.28C) and comprise -180° twists (FIG.28D). [00200] These mixer designs (Design Nos. A1, D10, D7, D6) and a control 0.5 mm cross mixer were tested in a simulation experiment to test total pressure and shear stress, as shown in Table 12. Table 12 [00201] Design No. D7 resulted in low total pressure and shear stress. These values were both lower than the original static mixer design (Design No. A1) simulation values. Design No. D7 also had a mixing efficiency 124% higher than the Koflo analogue Design No. A1. The batch sizes were approximately 8 times higher (both maximum and minimum) on the designed mixers as compared to the 0.5 mm cross mixer, allowing for high flow rates of approximately 905 mL/min total flow rate. [00202] A complete mixer design summary is shown in Table 13. The static mixer designs corresponding to those described in Table 13 are shown in FIGs.29A-C. Atty Dkt No.: 58530-742601 Table 13 Atty Dkt No.: 58530-742601 [00203] In summarizing Table 13, all of the above design conditions were evaluated and compared with a goal of minimizing both back-pressure and shear stress experienced by the reagents, while maximizing the speed of mixing of the reagents. A cutoff back-pressure of 10 psig was used to narrow down results, as this is 2/3 of the maximum rated pressure of a low- shear pump system we hope to use to drive the mixer flow. This cutoff was used to account for pressure increases from extra tubing as well as possible unquantified inaccuracy of the CFD results. Of the designs with low back-pressure, Design No. D7 had the highest simulated mixing speed as well as a low max average shear stress, making it a lead candidate. Design No. D6 was also considered due to its similar design to Design No. D7, but with almost 2 psi lower simulated back-pressure. Design No. D10 was considered due to its identical mixer design to Design No. D7 but with two inlets rather than four, thereby simplifying upstream connections. These three designs, along with a Koflo analogue, Design No. A1 were successfully manufactured and tested. Of these four manufactured designs, Design No. D7 performed best at lower flow rates and was chosen to be tested at the higher simulated total flow rate of ~905 mL/min. At this speed, the back-pressure was measured to be approximately 15 psi, which is significantly higher than the simulated 6 psi back-pressure, but this was expected to a degree since the simulated back-pressure did not include contributions from upstream or downstream tubing and connections. Example 4: Manufactured Static Mixer Initial Tests [00204] Each of the selected manufactured static mixers were tested against a cross mixer and a tee mixer, all at a 3:1 buffer to lipid ratio (empties). A higher buffer pH can decrease particle size, so citrate buffer from pH 4 to pH 6 were tested. Buffer strength may also have an effect on particle size, so citrate buffer strength will be varied as well as adding sodium chloride (NaCl) to change charge screening and encourage particle formation. All conditions and tests performed are shown in Table 14 for empty LNPs with a FRR (Lipid:mRNA) ratio of 1:3. Table 14 Atty Dkt No.: 58530-742601 *Reynolds numbers based on initial SM control rates [00205] The initial tests (Tests 1-7) and control comparison results are shown in Table 15. Table 15 Atty Dkt No.: 58530-742601 [00206] Of these results, cross and tee mixers resulted in lowest particle size (PS) and polydispersity index (PDI) compared to static mixers. The best-performing static mixer designs were Design No. A1 and Design No. D7. [00207] A follow-up test using Design No. D7 was done under different buffer compositions, as it performed best compared to the other non-Koflo designs. The results are shown in Table 16. Table 16 [00208] From the results and tables, Design No. D7 produced the best results out of the non- Koflo-equivalent static mixer designs with a post-PD10 particle size of 88 nm. Buffer strength, pH, and added NaCl did not show much effect on particle size, with a variation of only ~10 nm between conditions. In these experiments, Design No. D7 were ran at a quarter of the total flow rate it was designed to run at. Tests to run the static mixers at higher flow rate may increase formulation performance. Example 5: Manufactured Static Mixer High Flow Tests on Peristaltic Pumps [00209] High flow rate tests were performed on Design No. D7 mixer to reach the simulated total flow rate of 904.8 mL/min. Design No. D7 had the most simulated mixing efficiency of the designs simulated at 904.8 mL/min total flow rate (TFR), and compared to Designs Nos. D10, D6, Design No. D7 produced smaller particle sizes at lower flow rates. Atty Dkt No.: 58530-742601 [00210] Peristaltic pumps mixing was tested through Design No. D7 with 3:1 flow rate ratio (FRR) of water/ethanol to measure pressures. Peristaltic pumps were chosen for the ease of reaching scaled-up flow rates. As the results show in Table 17 for testing at different total flow rates, at 904.8 mL/min total flow rate, the mRNA pump pressure was 15.4 psi, so these flow rates may be feasible with Levitronix pumps. Table 17 [00211] Tests were performed with RTX0051-Empty formulation at different total flow rates, as shown in Table 18. As the results show in Table 18, only 395.9 mL/min total flow rate produced 80 nm particles. Other total flow rates produced 106-119 nm particles post-PD10. It is postulated that that the decrease in particle size at 395.6 mL/min is due to the pulsation at 396 mL/min rather than it being the effect of total flow rates higher than 395.6 mL/min. Table 18 [00212] The results shown Design No. D7 mixer was able to withstand total flow rates of up to 904.8 mL/min, with a max pressure of only 15.4 psig (mRNA Pressure (psig)). This was considered to be very low pressure in view of the flow rates. Particle size was high at higher flow rates, with smallest particle size at the lowest flow rate tested (395.9 mL/min). The particle size Atty Dkt No.: 58530-742601 at this flow rate was ~86nm post-dilution. It was postulated that the pulsation from the peristaltic mixers at high flow conditions were causing non-ideal mixing conditions. Example 6: Manufactured Static Mixer High Flow Tests on HPLC Pumps [00213] High performance liquid chromatography (HPLC) pumps were used to see if it will create lower-sized particles at high flow rates as compared to particles generated from peristaltic pumps. Knauer BlueShadow 80P LC pumps (1000 mL/min) was used to achieve the required flow rates, and different parameters as were tested on peristaltic pumps. The simulated flow parameters were 904.8 total flow rate (mL/min), max back-pressure of 5.77 psig, and a mixing efficiency of 224% as compared to the original static design. Table 19 shows the test conditions on the HPLC pumps, which were set to mimic the test conditions on the peristaltic pump. Table 19 [00214] Tests were performed with RTX0051-Empty formulation at different total flow rates. As shown in Table 20, the four tested conditions resulted in smaller particles than those made with the peristaltic setup. The particle size ranged from 60-75 nm post-PD-10, with the highest flow condition resulting in the best particles across all sampling regimes. Minimal trend could be seen with regards to total flow rate, with all conditions creating decent empty particles. Table 20 Atty Dkt No.: 58530-742601 [00215] As the results indicate, the HPLC pumps generated particles were much smaller. Peristaltic fluctuations may have a major effect on mixing efficiency at high flow rates. With the Design No. D7 mixer, sub-65 nm empty particles were achieved as shown in Table 20. This demonstrated a 20 nm reduction in size compared to the Koflo analogue, Design No. A1, which generated ~84 to 87 nm particles as shown in Table 15. [00216] While preferred embodiments of the present disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. It is not intended that the disclosure be limited by the specific examples provided within the specification. While the disclosure has been described with reference to the aforementioned specification, the descriptions and illustrations of the embodiments herein are not meant to be construed in a limiting sense. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the disclosure. Furthermore, it shall be understood that all aspects of the disclosure are not limited to the specific depictions, configurations or relative proportions set forth herein which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the disclosure described herein may be employed in practicing the disclosure. It is therefore contemplated that the disclosure shall also cover any such alternatives, modifications, variations or equivalents. It is intended that the following claims define the scope of the disclosure and that methods and structures within the scope of these claims and their equivalents be covered thereby. EMBODIMENTS [00217] The following are exemplary embodiments of the disclosure herein: Embodiment 1. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; Atty Dkt No.: 58530-742601 iii. an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is no greater than about 0.25. Embodiment 2. The system of embodiment 1, wherein the conduit is tee-shaped. Embodiment 3. The system of embodiment 1, wherein the conduit is Y-shaped. Embodiment 4. The system of embodiment 3, wherein the conduit further comprises a cut zip tie. Embodiment 5. The system of embodiment 1, wherein the conduit is L-shaped. Embodiment 6. The system of any one of the preceding embodiments, wherein the static mixer is less than about 40 millimeters (mm) in length. Embodiment 7. The system of any one of the preceding embodiments, wherein the static mixer is less than about 10 mm in length. Embodiment 8. The system of any one of the preceding embodiments, wherein the static mixer is less than about 5 mm in length. Embodiment 9. The system of any one of the preceding embodiments, wherein the static mixer is about 4.8 mm in length. Embodiment 10. The system of any one of the preceding embodiments, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 11. The system of embodiment 10, wherein the inner diameter is about ¼ inch. Embodiment 12. The system of embodiment 10, wherein the inner diameter is about 1/8 inch. Embodiment 13. The system of any one of the preceding embodiments, wherein the polydispersity index is about 0.05 to about 0.25. Embodiment 14. The system of embodiment 13, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 15. The system of any one of embodiments 1-12, wherein the polydispersity index is no greater than about 0.1. Embodiment 16. The system of any one of the preceding embodiments, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Atty Dkt No.: 58530-742601 Embodiment 17. The system of embodiment 16, wherein the Reynolds number is about 500 to about 1000. Embodiment 18. The system of embodiment 16, wherein the Reynolds number is about 1000. Embodiment 19. The system of any one of the preceding embodiments, wherein the lipid nanoparticle is no greater than about 100 nanometers (nm). Embodiment 20. The system of embodiment 19, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 21. The system of embodiment 19, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 22. The system of any one of the preceding embodiments, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 23. The system of embodiment 22, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 24. The system of embodiment 22, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 25. The system of any one of the preceding embodiments, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 26. The system of any one of the preceding embodiments, wherein the static mixer comprises 8 or more elements. Embodiment 27. The system of any one of the preceding embodiments, wherein the static mixer comprises 12 or more elements. Embodiment 28. The system of any one of the preceding embodiments, wherein the static mixer comprises 16 or more elements. Embodiment 29. The system of any one of the preceding embodiments, wherein the static mixer comprises 24 or more elements. Embodiment 30. The system of any one of the preceding embodiments, wherein the static mixer is downstream of the conduit. Embodiment 31. A method of generating a lipid nanoparticle, the method comprising: (a) providing: Atty Dkt No.: 58530-742601 i. a conduit comprising: a. a first inlet; b. a second inlet; c. an outlet; and ii. a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid, wherein a polydispersity index of the mixture fluid is no greater than about 0.25. Embodiment 32. The method of embodiment 31, wherein the conduit is tee-shaped. Embodiment 33. The method of embodiment 31, wherein the conduit is Y-shaped. Embodiment 34. The method of embodiment 33, wherein the conduit further comprises a cut zip tie. Embodiment 35. The method of embodiment 31, wherein the conduit is L-shaped. Embodiment 36. The method of any one of embodiments 31-35, wherein the static mixer is less than about 40 mm in length. Embodiment 37. The method of any one of embodiments 31-36, wherein the static mixer is less than about 10 mm in length. Embodiment 38. The method of any one of embodiments 31-37, wherein the static mixer is less than about 5 mm in length. Embodiment 39. The method of any one of embodiments 31-38, wherein the static mixer is about 4.8 mm in length. Embodiment 40. The method of any one of embodiments 31-39, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 41. The method of embodiment 40, wherein the inner diameter is about ¼ inch. Embodiment 42. The method of embodiment 40, wherein the inner diameter is about 1/8 inch. Embodiment 43. The method of any one of embodiments 31-42, wherein the polydispersity index is about 0.05 to about 0.25. Atty Dkt No.: 58530-742601 Embodiment 44. The method of embodiment 43, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 45. The method of any one of embodiments 31-42, wherein the polydispersity index is no greater than about 0.1. Embodiment 46. The method of any one of embodiments 31-45, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 47. The method of embodiment 46, wherein the Reynolds number is about 500 to about 1000. Embodiment 48. The method of embodiment 46, wherein the Reynolds number is about 1000. Embodiment 49. The method of any one of embodiments 31-48, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 50. The method of embodiment 49, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 51. The method of embodiment 49, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 52. The method of any one of embodiments 31-51, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 53. The method of embodiment 52, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 54. The method of embodiment 52, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 55. The method of any one of embodiments 31-54, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 56. The method of any one of embodiments 31-55, wherein the static mixer comprises 8 or more elements. Embodiment 57. The method of any one of embodiments 31-56, wherein the static mixer comprises 12 or more elements. Embodiment 58. The method of any one of embodiments 31-57, wherein the static mixer comprises 16 or more elements. Atty Dkt No.: 58530-742601 Embodiment 59. The method of any one of embodiments 31-58, wherein the static mixer comprises 24 or more elements. Embodiment 60. The method of any one of embodiments 31-59, wherein the static mixer is downstream of the conduit. Embodiment 61. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; iii. a junction site wherein the first inlet and the second inlet intersects; and (b) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site. Embodiment 62. The system of embodiment 61, wherein the conduit is tee-shaped. Embodiment 63. The system of embodiment 61, wherein the conduit is Y-shaped. Embodiment 64. The system of embodiment 63, wherein the conduit further comprises a cut zip tie. Embodiment 65. The system of embodiment 61, wherein the conduit is L-shaped. Embodiment 66. The system of any one of embodiments 61-65, wherein the static mixer is less than about 40 mm in length. Embodiment 67. The system of any one of embodiments 61-66, wherein the static mixer is less than about 10 mm in length. Embodiment 68. The system of any one of embodiments 61-67, wherein the static mixer is less than about 5 mm in length. Embodiment 69. The system of any one of embodiments 61-68, wherein the static mixer is about 4.8 mm in length. Embodiment 70. The system of any one of embodiments 61-69, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 71. The system of embodiment 70, wherein the inner diameter is about ¼ inch. Embodiment 72. The system of embodiment 70, wherein the inner diameter is about 1/8 inch. Atty Dkt No.: 58530-742601 Embodiment 73. The system of any one of embodiments 61-72, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 74. The system of embodiment 73, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 75. The system of any one of embodiments 61-72, wherein the polydispersity index is no greater than about 0.1. Embodiment 76. The system of any one of embodiments 61-75, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 77. The system of embodiment 76, wherein the Reynolds number is about 500 to about 1000. Embodiment 78. The system of embodiment 76, wherein the Reynolds number is about 1000. Embodiment 79. The system of any one of embodiments 61-78, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 80. The system of embodiment 79, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 81. The system of embodiment 79, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 82. The system of any one of embodiments 61-81, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 83. The system of embodiment 82, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 84. The system of embodiment 82, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 85. The system of any one of embodiments 61-84, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 86. The system of any one of embodiments 61-85, wherein the static mixer comprises 8 or more elements. Embodiment 87. The system of any one of embodiments 61-86, wherein the static mixer comprises 12 or more elements. Atty Dkt No.: 58530-742601 Embodiment 88. The system of any one of embodiments 61-87, wherein the static mixer comprises 16 or more elements. Embodiment 89. The system of any one of embodiments 61-88, wherein the static mixer comprises 24 or more elements. Embodiment 90. The system of any one of embodiments 61-89, wherein the static mixer is downstream of the conduit. Embodiment 91. A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit comprising: a. a first inlet; b. a second inlet; c. a junction site downstream of the first inlet and the second inlet; and ii. a static mixer, wherein the static mixer is disposed at a distal end of the junction site; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the junction site; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site. Embodiment 92. The method of embodiment 91, wherein the conduit is tee-shaped. Embodiment 93. The method of embodiment 91, wherein the conduit is Y-shaped. Embodiment 94. The method of embodiment 93, wherein the conduit further comprises a cut zip tie. Embodiment 95. The method of embodiment 91, wherein the conduit is L-shaped. Embodiment 96. The method of any one of embodiments 91-95, wherein the static mixer is less than about 40 mm in length. Embodiment 97. The method of any one of embodiments 91-96, wherein the static mixer is less than about 10 mm in length. Embodiment 98. The method of any one of embodiments 91-97, wherein the static mixer is less than about 5 mm in length. Embodiment 99. The method of any one of embodiments 91-98, wherein the static mixer is about 4.8 mm in length. Atty Dkt No.: 58530-742601 Embodiment 100. The method of any one of embodiments 91-99, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 101. The method of embodiment 100, wherein the inner diameter is about ¼ inch. Embodiment 102. The method of embodiment 100, wherein the inner diameter is about 1/8 inch. Embodiment 103. The method of any one of embodiments 91-102, wherein the nucleic acid and the one or more lipid components of I are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 104. The method of embodiment 103, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 105. The method of any one of embodiments 91-102, wherein the polydispersity index is no greater than about 0.1. Embodiment 106. The method of any one of embodiments 91-105, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 107. The method of embodiment 106, wherein the Reynolds number is about 500 to about 1000. Embodiment 108. The method of embodiment 106, wherein the Reynolds number is about 1000. Embodiment 109. The method of any one of embodiments 91-108, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 110. The method of embodiment 109, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 111. The method of embodiment 109, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 112. The method of any one of embodiments 91-111, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 113. The method of embodiment 112, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 114. The method of embodiment 112, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 115. The method of any one of embodiments 91-114, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby Atty Dkt No.: 58530-742601 encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 116. The method of any one of embodiments 91-115, wherein the static mixer comprises 8 or more elements. Embodiment 117. The method of any one of embodiments 91-116, wherein the static mixer comprises 12 or more elements. Embodiment 118. The method of any one of embodiments 91-117, wherein the static mixer comprises 16 or more elements. Embodiment 119. The method of any one of embodiments 91-118, wherein the static mixer comprises 24 or more elements. Embodiment 120. The method of any one of embodiments 91-119, wherein the static mixer is downstream of the conduit. Embodiment 121. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; iii. an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a Reynolds number of the mixture fluid is no greater than about 1000. Embodiment 122. The system of embodiment 121, wherein the conduit is tee-shaped. Embodiment 123. The system of embodiment 121, wherein the conduit is Y-shaped. Embodiment 124. The system of embodiment 123, wherein the conduit further comprises a cut zip tie. Embodiment 125. The system of embodiment 121, wherein the conduit is L-shaped. Embodiment 126. The system of any one of embodiments 121-125, wherein the static mixer is less than about 40 mm in length. Embodiment 127. The system of any one of embodiments 121-126, wherein the static mixer is less than about 10 mm in length. Embodiment 128. The system of any one of embodiments 121-127, wherein the static mixer is less than about 5 mm in length. Atty Dkt No.: 58530-742601 Embodiment 129. The system of any one of embodiments 121-128, wherein the static mixer is about 4.8 mm in length. Embodiment 130. The system of any one of embodiments 121-129, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 131. The system of embodiment 130, wherein the inner diameter is about ¼ inch. Embodiment 132. The system of embodiment 130, wherein the inner diameter is about 1/8 inch. Embodiment 133. The system of any one of embodiments 121-132, wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 134. The system of embodiment 133, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 135. The system of any one of embodiments 121-132, wherein the polydispersity index is no greater than about 0.1. Embodiment 136. The system of any one of embodiments 121-135, wherein the Reynolds number is about 150 to about 1000. Embodiment 137. The system of embodiment 136, wherein the Reynolds number is about 500 to about 1000. Embodiment 138. The system of embodiment 136, wherein the Reynolds number is about 1000. Embodiment 139. The system of any one of embodiments 121-138, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 140. The system of embodiment 139, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 141. The system of embodiment 139, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 142. The system of any one of embodiments 121-141, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 143. The system of embodiment 142, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 144. The system of embodiment 142, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Atty Dkt No.: 58530-742601 Embodiment 145. The system of any one of embodiments 121-144, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 146. The system of any one of embodiments 121-145, wherein the static mixer comprises 8 or more elements. Embodiment 147. The system of any one of embodiments 121-146, wherein the static mixer comprises 12 or more elements. Embodiment 148. The system of any one of embodiments 121-147, wherein the static mixer comprises 16 or more elements. Embodiment 149. The system of any one of embodiments 121-148, wherein the static mixer comprises 24 or more elements. Embodiment 150. The system of any one of embodiments 121-149, wherein the static mixer is downstream of the conduit. Embodiment 151. A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit comprising: a. a first inlet; b. a second inlet; c. an outlet; and ii. a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid, wherein a Reynolds number of the mixture fluid is no greater than about 1000. Embodiment 152. The method of embodiment 151, wherein the conduit is tee-shaped. Embodiment 153. The method of embodiment 151, wherein the conduit is Y-shaped. Embodiment 154. The method of embodiment 153, wherein the conduit further comprises a cut zip tie. Embodiment 155. The method of embodiment 151, wherein the conduit is L-shaped. Atty Dkt No.: 58530-742601 Embodiment 156. The method of any one of embodiments 151-155, wherein the static mixer is less than about 40 mm in length. Embodiment 157. The method of any one of embodiments 151-156, wherein the static mixer is less than about 10 mm in length. Embodiment 158. The method of any one of embodiments 151-157, wherein the static mixer is less than about 5 mm in length. Embodiment 159. The method of any one of embodiments 151-158, wherein the static mixer is about 4.8 mm in length. Embodiment 160. The method of any one of embodiments 151-159, wherein the static mixer has an inner diameter of about 1/16 inch to abo½1/2 inch. Embodiment 161. The method of embodiment 160, wherein the inner diameter is abo¼1/4 inch. Embodiment 162. The method of embodiment 160, wherein the inner diameter is about 1/8 inch. Embodiment 163. The method of any one of embodiments 151-162, wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 164. The method of embodiment 163, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 165. The method of any one of embodiments 151-162, wherein the polydispersity index is no greater than about 0.1. Embodiment 166. The method of any one of embodiments 151-165, wherein the Reynolds number is about 150 to about 1000. Embodiment 167. The method of embodiment 166, wherein the Reynolds number is about 500 to about 1000. Embodiment 168. The method of embodiment 166, wherein the Reynolds number is about 1000. Embodiment 169. The method of any one of embodiments 151-168, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 170. The method of embodiment 169, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 171. The method of embodiment 169, wherein the lipid nanoparticle is about no greater than about 50 nm. Atty Dkt No.: 58530-742601 Embodiment 172. The method of any one of embodiments 151-171, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 173. The method of embodiment 172, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 174. The method of embodiment 172, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 175. The method of any one of embodiments 151-174, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 176. The method of any one of embodiments 151-175, wherein the static mixer comprises 8 or more elements. Embodiment 177. The method of any one of embodiments 151-176, wherein the static mixer comprises 12 or more elements. Embodiment 178. The method of any one of embodiments 151-177, wherein the static mixer comprises 16 or more elements. Embodiment 179. The method of any one of embodiments 151-178, wherein the static mixer comprises 24 or more elements. Embodiment 180. The method of any one of embodiments 151-179, wherein the static mixer is downstream of the conduit. Embodiment 181. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; iii. an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min. Embodiment 182. The system of embodiment 181, wherein the conduit is tee-shaped. Embodiment 183. The system of embodiment 181, wherein the conduit is Y-shaped. Embodiment 184. The system of embodiment 183, wherein the conduit further comprises a cut zip tie. Atty Dkt No.: 58530-742601 Embodiment 185. The system of embodiment 181, wherein the conduit is L-shaped. Embodiment 186. The system of any one of embodiments 181-185, wherein the static mixer is less than about 40 mm in length. Embodiment 187. The system of any one of embodiments 181-186, wherein the static mixer is less than about 10 mm in length. Embodiment 188. The system of any one of embodiments 181-187, wherein the static mixer is less than about 5 mm in length. Embodiment 189. The system of any one of embodiments 181-188, wherein the static mixer is about 4.8 mm in length. Embodiment 190. The system of any one of embodiments 181-189, wherein the static mixer has an inner diameter of about 1/16 inch to abo½1/2 inch. Embodiment 191. The system of embodiment 190, wherein the inner diameter is about ¼ inch. Embodiment 192. The system of embodiment 190, wherein the inner diameter is about 1/8 inch. Embodiment 193. The system of any one of embodiments 181-192, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 194. The system of embodiment 193, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 195. The system of any one of embodiments 181-192, wherein the polydispersity index is no greater than about 0.1. Embodiment 196. The system of any one of embodiments 181-195, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 197. The system of embodiment 196, wherein the Reynolds number is about 500 to about 1000. Embodiment 198. The system of embodiment 196, wherein the Reynolds number is about 1000. Embodiment 199. The system of any one of embodiments 181-198, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 200. The system of embodiment 199, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Atty Dkt No.: 58530-742601 Embodiment 201. The system of embodiment 199, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 202. The system of any one of embodiments 181-201, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 203. The system of embodiment 202, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 204. The system of embodiment 202, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 205. The system of any one of embodiments 181-204, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 206. The system of any one of embodiments 181-205, wherein the static mixer comprises 8 or more elements. Embodiment 207. The system of any one of embodiments 181-206, wherein the static mixer comprises 12 or more elements. Embodiment 208. The system of any one of embodiments 181-207, wherein the static mixer comprises 16 or more elements. Embodiment 209. The system of any one of embodiments 181-208, wherein the static mixer comprises 24 or more elements. Embodiment 210. The system of any one of embodiments 181-209, wherein the static mixer is downstream of the conduit. Embodiment 211. A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit comprising: a. a first inlet; b. a second inlet; c. an outlet; and ii. a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and Atty Dkt No.: 58530-742601 (e) using the static mixer to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min. Embodiment 212. The method of embodiment 211, wherein the conduit is tee-shaped. Embodiment 213. The method of embodiment 211, wherein the conduit is Y-shaped. Embodiment 214. The method of embodiment 213, wherein the conduit further comprises a cut zip tie. Embodiment 215. The method of embodiment 211, wherein the conduit is L-shaped. Embodiment 216. The method of any one of embodiments 211-215, wherein the static mixer is less than about 40 mm in length. Embodiment 217. The method of any one of embodiments 211-216, wherein the static mixer is less than about 10 mm in length. Embodiment 218. The method of any one of embodiments 211-217, wherein the static mixer is less than about 5 mm in length. Embodiment 219. The method of any one of embodiments 211-218, wherein the static mixer is about 4.8 mm in length. Embodiment 220. The method of any one of embodiments 211-219, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 221. The method of embodiment 220, wherein the inner diameter is about ¼ inch. Embodiment 222. The method of embodiment 220, wherein the inner diameter is about 1/8 inch. Embodiment 223. The method of any one of embodiments 211-222, wherein the nucleic acid and the one or more lipid components of € are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 224. The method of embodiment 223, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 225. The method of any one of embodiments 211-222, wherein the polydispersity index is no greater than about 0.1. Embodiment 226. The method of any one of embodiments 211-225, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 227. The method of embodiment 226, wherein the Reynolds number is about 500 to about 1000. Atty Dkt No.: 58530-742601 Embodiment 228. The method of embodiment 226, wherein the Reynolds number is about 1000. Embodiment 229. The method of any one of embodiments 211-228, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 230. The method of embodiment 229, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 231. The method of embodiment 229, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 232. The method of any one of embodiments 211-231, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 233. The method of embodiment 232, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 234. The method of embodiment 232, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 235. The method of any one of embodiments 211-234, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 236. The method of any one of embodiments 211-235, wherein the static mixer comprises 8 or more elements. Embodiment 237. The method of any one of embodiments 211-236, wherein the static mixer comprises 12 or more elements. Embodiment 238. The method of any one of embodiments 211-237, wherein the static mixer comprises 16 or more elements. Embodiment 239. The method of any one of embodiments 211-238, wherein the static mixer comprises 24 or more elements. Embodiment 240. The method of any one of embodiments 211-239, wherein the static mixer is downstream of the conduit. Embodiment 241. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; Atty Dkt No.: 58530-742601 iii. an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the lipid nanoparticle is no greater than about 150 nm. Embodiment 242. The system of embodiment 241, wherein the conduit is tee-shaped. Embodiment 243. The system of embodiment 241, wherein the conduit is Y-shaped. Embodiment 244. The system of embodiment 243, wherein the conduit further comprises a cut zip tie. Embodiment 245. The system of embodiment 241, wherein the conduit is L-shaped. Embodiment 246. The system of any one of embodiments 241-245, wherein the static mixer is less than about 40 mm in length. Embodiment 247. The system of any one of embodiments 241-246, wherein the static mixer is less than about 10 mm in length. Embodiment 248. The system of any one of embodiments 241-247, wherein the static mixer is less than about 5 mm in length. Embodiment 249. The system of any one of embodiments 241-248, wherein the static mixer is about 4.8 mm in length. Embodiment 250. The system of any one of embodiments 241-249, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 251. The system of embodiment 250, wherein the inner diameter is about ¼ inch. Embodiment 252. The system of embodiment 250, wherein the inner diameter is about 1/8 inch. Embodiment 253. The system of any one of embodiments 241-252, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 254. The system of embodiment 253, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 255. The system of any one of embodiments 241-252, wherein the polydispersity index is no greater than about 0.1. Embodiment 256. The system of any one of embodiments 241-255, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Atty Dkt No.: 58530-742601 Embodiment 257. The system of embodiment 256, wherein the Reynolds number is about 500 to about 1000. Embodiment 258. The system of embodiment 256, wherein the Reynolds number is about 1000. Embodiment 259. The system of any one of embodiments 241-258, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 260. The system of embodiment 259, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 261. The system of embodiment 259, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 262. The system of any one of embodiments 241-261, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 263. The system of embodiment 262, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 264. The system of embodiment 262, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 265. The system of any one of embodiments 241-264, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 266. The system of any one of embodiments 241-265, wherein the static mixer comprises 8 or more elements. Embodiment 267. The system of any one of embodiments 241-266, wherein the static mixer comprises 12 or more elements. Embodiment 268. The system of any one of embodiments 241-267, wherein the static mixer comprises 16 or more elements. Embodiment 269. The system of any one of embodiments 241-268, wherein the static mixer comprises 24 or more elements. Embodiment 270. The system of any one of embodiments 241-269, wherein the static mixer is downstream of the conduit. Embodiment 271. A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit comprising: Atty Dkt No.: 58530-742601 a. a first inlet; b. a second inlet; c. an outlet; and ii. a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the lipid nanoparticle is no greater than about 150 nm. Embodiment 272. The method of embodiment 271, wherein the conduit is tee-shaped. Embodiment 273. The method of embodiment 271, wherein the conduit is Y-shaped. Embodiment 274. The method of embodiment 273, wherein the conduit further comprises a cut zip tie. Embodiment 275. The method of embodiment 271, wherein the conduit is L-shaped. Embodiment 276. The method of any one of embodiments 271-275, wherein the static mixer is less than about 40 mm in length. Embodiment 277. The method of any one of embodiments 271-276, wherein the static mixer is less than about 10 mm in length. Embodiment 278. The method of any one of embodiments 271-277, wherein the static mixer is less than about 5 mm in length. Embodiment 279. The method of any one of embodiments 271-278, wherein the static mixer is about 4.8 mm in length. Embodiment 280. The method of any one of embodiments 271-279, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 281. The method of embodiment 280, wherein the inner diameter is about ¼ inch. Embodiment 282. The method of embodiment 280, wherein the inner diameter is about 1/8 inch. Embodiment 283. The method of any one of embodiments 271-282, wherein the nucleic acid and the one or more lipid components of € are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 284. The method of embodiment 283, wherein the polydispersity index is about 0.1 to about 0.2. Atty Dkt No.: 58530-742601 Embodiment 285. The method of any one of embodiments 271-282, wherein the polydispersity index is no greater than about 0.1. Embodiment 286. The method of any one of embodiments 271-285, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 287. The method of embodiment 286, wherein the Reynolds number is about 500 to about 1000. Embodiment 288. The method of embodiment 286, wherein the Reynolds number is about 1000. Embodiment 289. The method of any one of embodiments 271-288, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 290. The method of embodiment 289, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 291. The method of embodiment 289, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 292. The method of any one of embodiments 271-291, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 293. The method of embodiment 292, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 294. The method of embodiment 292, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 295. The method of any one of embodiments 271-294, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 296. The method of any one of embodiments 271-295, wherein the static mixer comprises 8 or more elements. Embodiment 297. The method of any one of embodiments 271-296, wherein the static mixer comprises 12 or more elements. Embodiment 298. The method of any one of embodiments 271-297, wherein the static mixer comprises 16 or more elements. Embodiment 299. The method of any one of embodiments 271-298, wherein the static mixer comprises 24 or more elements. Atty Dkt No.: 58530-742601 Embodiment 300. The method of any one of embodiments 271-299, wherein the static mixer is downstream of the conduit. Embodiment 301. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit, comprising: i. a first inlet configured to introduce a nucleic acid; ii. a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; iii. an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer configured to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 302. The system of embodiment 301, wherein the conduit is tee-shaped. Embodiment 303. The system of embodiment 301, wherein the conduit is Y-shaped. Embodiment 304. The system of embodiment 303, wherein the conduit further comprises a cut zip tie. Embodiment 305. The system of embodiment 301, wherein the conduit is L-shaped. Embodiment 306. The system of any one of embodiments 301-305, wherein the static mixer is less than about 40 mm in length. Embodiment 307. The system of any one of embodiments 301-306, wherein the static mixer is less than about 10 mm in length. Embodiment 308. The system of any one of embodiments 301-307, wherein the static mixer is less than about 5 mm in length. Embodiment 309. The system of any one of embodiments 301-308, wherein the static mixer is about 4.8 mm in length. Embodiment 310. The system of any one of embodiments 301-309, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 311. The system of embodiment 310, wherein the inner diameter is about ¼ inch. Embodiment 312. The system of embodiment 310, wherein the inner diameter is about 1/8 inch. Atty Dkt No.: 58530-742601 Embodiment 313. The system of any one of embodiments 301-312, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 314. The system of embodiment 313, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 315. The system of any one of embodiments 301-312, wherein the polydispersity index is no greater than about 0.1. Embodiment 316. The system of any one of embodiments 301-315, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 317. The system of embodiment 316, wherein the Reynolds number is about 500 to about 1000. Embodiment 318. The system of embodiment 316, wherein the Reynolds number is about 1000. Embodiment 319. The system of any one of embodiments 301-318, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 320. The system of embodiment 319, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 321. The system of embodiment 319, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 322. The system of any one of embodiments 301-321, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 323. The system of embodiment 322, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 324. The system of embodiment 322, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 325. The system of any one of embodiments 301-324, wherein the static mixer comprises 8 or more elements. Embodiment 326. The system of any one of embodiments 301-325, wherein the static mixer comprises 12 or more elements. Embodiment 327. The system of any one of embodiments 301-326, wherein the static mixer comprises 16 or more elements. Embodiment 328. The system of any one of embodiments 301-327, wherein the static mixer comprises 24 or more elements. Atty Dkt No.: 58530-742601 Embodiment 329. The system of any one of embodiments 301-328, wherein the static mixer is downstream of the conduit. Embodiment 330. A method of generating a lipid nanoparticle, the method comprising: (a) providing: i. a conduit, comprising: a. a first inlet; b. a second inlet; c. an outlet; and ii. a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 331. The method of embodiment 330, wherein the conduit is tee-shaped. Embodiment 332. The method of embodiment 330, wherein the conduit is Y-shaped. Embodiment 333. The method of embodiment 332, wherein the conduit further comprises a cut zip tie. Embodiment 334. The method of embodiment 330, wherein the conduit is L-shaped. Embodiment 335. The method of any one of embodiments 330-334, wherein the static mixer is less than about 40 mm in length. Embodiment 336. The method of any one of embodiments 330-335, wherein the static mixer is less than about 10 mm in length. Embodiment 337. The method of any one of embodiments 330-336, wherein the static mixer is less than about 5 mm in length. Embodiment 338. The method of any one of embodiments 330-337, wherein the static mixer is about 4.8 mm in length. Embodiment 339. The method of any one of embodiments 330-338, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 340. The method of embodiment 339, wherein the inner diameter is about ¼ inch. Atty Dkt No.: 58530-742601 Embodiment 341. The method of embodiment 339, wherein the inner diameter is about 1/8 inch. Embodiment 342. The method of any one of embodiments 330-341, wherein the nucleic acid and the one or more lipid components of € are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 343. The method of embodiment 342, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 344. The method of any one of embodiments 330-341, wherein the polydispersity index is no greater than about 0.1. Embodiment 345. The method of any one of embodiments 330-344, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 346. The method of embodiment 345, wherein the Reynolds number is about 500 to about 1000. Embodiment 347. The method of embodiment 345, wherein the Reynolds number is about 1000. Embodiment 348. The method of any one of embodiments 330-347, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 349. The method of embodiment 348, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 350. The method of embodiment 348, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 351. The method of any one of embodiments 330-350, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 352. The method of embodiment 351, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 353. The method of embodiment 351, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 354. The method of any one of embodiments 330-353, wherein the static mixer comprises 8 or more elements. Embodiment 355. The method of any one of embodiments 330-354, wherein the static mixer comprises 12 or more elements. Embodiment 356. The method of any one of embodiments 330-355, wherein the static mixer comprises 16 or more elements. Atty Dkt No.: 58530-742601 Embodiment 357. The method of any one of embodiments 330-356, wherein the static mixer comprises 24 or more elements. Embodiment 358. The method of any one of embodiments 330-357, wherein the static mixer is downstream of the conduit. Embodiment 359. A system for generating a lipid nanoparticle, the system comprising a static mixer, wherein when a nucleic acid and one or more lipid components are joined in the static mixer, the system reaches a pressure of no greater than 8 psig. Embodiment 360. The system of embodiment 359, wherein the system further comprises a first inlet configured to introduce a nucleic acid. Embodiment 361. The system of embodiment 359 or 360, wherein the system further comprises a second inlet configured to introduce one or more lipid components of the lipid nanoparticle. Embodiment 362. The system of any one of embodiments 359-361, wherein the static mixer is less than about 40 mm in length. Embodiment 363. The system of any one of embodiments 359-362, wherein the static mixer is less than about 10 mm in length. Embodiment 364. The system of any one of embodiments 359-363, wherein the static mixer is less than about 5 mm in length. Embodiment 365. The system of any one of embodiments 359-364, wherein the static mixer is about 4.8 mm in length. Embodiment 366. The system of any one of embodiments 359-365, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. Embodiment 367. The system of embodiment 366, wherein the inner diameter is about ¼ inch. Embodiment 368. The system of embodiment 366, wherein the inner diameter is about 1/8 inch. Embodiment 369. The system of any one of embodiments 359-368, wherein the nucleic acid and the one or more lipid components of (b) are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is about 0.05 to about 0.25. Embodiment 370. The system of embodiment 369, wherein the polydispersity index is about 0.1 to about 0.2. Embodiment 371. The system of any one of embodiments 369-370, wherein the polydispersity index is no greater than about 0.1. Atty Dkt No.: 58530-742601 Embodiment 372. The system of any one of embodiments 359-371, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. Embodiment 373. The system of embodiment 372, wherein the Reynolds number is about 500 to about 1000. Embodiment 374. The system of embodiment 372, wherein the Reynolds number is about 1000. Embodiment 375. The system of any one of embodiments 359-374, wherein the lipid nanoparticle is no greater than about 100 nm. Embodiment 376. The system of embodiment 375, wherein the lipid nanoparticle is about 50 nm to about 100 nm. Embodiment 377. The system of embodiment 375, wherein the lipid nanoparticle is about no greater than about 50 nm. Embodiment 378. The system of any one of embodiments 359-377, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. Embodiment 379. The system of embodiment 378, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. Embodiment 380. The system of embodiment 378, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. Embodiment 381. The system of any one of embodiments 359-380, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Embodiment 382. The system of any one of embodiments 359-381, wherein the static mixer comprises 8 or more elements. Embodiment 383. The system of any one of embodiments 359-382, wherein the static mixer comprises 12 or more elements. Embodiment 384. The system of any one of embodiments 359-383, wherein the static mixer comprises 16 or more elements. Embodiment 385. The system of any one of embodiments 359-384, wherein the static mixer comprises 24 or more elements.

Claims

Atty Dkt No.: 58530-742601 CLAIMS WHAT IS CLAIMED IS: 1. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit: (b) a static mixer, wherein a nucleic acid and one or more lipid components are mixed to generate a mixture fluid; and (c) a high performance liquid chromatography (HPLC) pump. 2. A system for generating a lipid nanoparticle, the system comprising a conduit and a static mixer, wherein a nucleic acid and one or more lipid components are mixed to generate a mixture fluid, and wherein when the nucleic acid and the one or more lipid components are joined in the static mixer, the system reaches a pressure of no greater than 8 pounds per square inch gauge (psig). 3. The system of claim 1 or 2, wherein the conduit comprises a first inlet configured to introduce the nucleic acid. 4. The system of any one of claims 1-3, wherein the conduit comprises a second inlet configured to introduce one or more lipid components of the lipid nanoparticle. 5. The system of any one of claims 1-4, wherein the conduit comprises an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet. 6. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: (i) a first inlet configured to introduce a nucleic acid; (ii) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; (iii) an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a polydispersity index of the mixture fluid is no greater than about 0.25. 7. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: (i) a first inlet configured to introduce a nucleic acid; (ii) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; Atty Dkt No.: 58530-742601 (iii) a junction site wherein the first inlet and the second inlet intersects; and (b) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site. 8. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: (i) a first inlet configured to introduce a nucleic acid; (ii) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; (iii) an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer, wherein the nucleic acid and the one or more lipid components are mixed to generate a mixture fluid, and wherein a Reynolds number of the mixture fluid is no greater than about 1000. 9. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: (i) a first inlet configured to introduce a nucleic acid; (ii) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; (iii) an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min. 10. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit comprising: (i) a first inlet configured to introduce a nucleic acid; (ii) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; (iii) an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the lipid nanoparticle is no greater than about 150 nm. Atty Dkt No.: 58530-742601 11. A system for generating a lipid nanoparticle, the system comprising: (a) a conduit, comprising: (i) a first inlet configured to introduce a nucleic acid; (ii) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; (iii) an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (b) a static mixer configured to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. 12. A system for generating a lipid nanoparticle, comprising: (i) a conduit comprising: (a) a first inlet configured to introduce a nucleic acid; (b) a second inlet configured to introduce one or more lipid components of the lipid nanoparticle; (c) an outlet, wherein the nucleic acid and the one or more lipid components are joined in the outlet; and (ii) a static mixer configured to mix the nucleic acid and the one or more lipid components, wherein when a nucleic acid and one or more lipid components are mixed in the static mixer, the system reaches a pressure of no greater than 8 pounds per square inch gauge (psig). 13. The system of any one of the preceding claims, wherein the conduit comprises a third inlet configured to introduce the nucleic acid or one or more lipid components of the lipid nanoparticle. 14. The system of any one of the preceding claims, wherein the conduit comprises a fourth inlet configured to introduce the nucleic acid or the one or more lipid components of the lipid nanoparticle. 15. The system of any one of the preceding claims, wherein the static mixer comprises a plurality of elements, wherein each element of the static mixer makes a 270° twist every 1/8 inch. 16. The system of any one of the preceding claims, wherein the system comprises a channel housing the static mixer configured to mix the nucleic acid and the one or more lipid components, wherein the channel is square shaped and makes a -180° twist every 1/8 inch. 17. The system of any one of the preceding claims, wherein the conduit is tee-shaped. Atty Dkt No.: 58530-742601 18. The system of any one of claims 1-17, wherein the conduit is Y-shaped. 19. The system of any one of claims 1-17, wherein the conduit is L-shaped. 20. The system of any one of the preceding claims, wherein the conduit further comprises a cut zip tie. 21. The system of any one of the preceding claims, wherein the static mixer is less than about 40 millimeters (mm) in length. 22. The system of any one of the preceding claims, wherein the static mixer is less than about 10 mm in length. 23. The system of any one of the preceding claims, wherein the static mixer is less than about 5 mm in length. 24. The system of any one of the preceding claims, wherein the static mixer is about 4.8 mm in length. 25. The system of any one of the preceding claims, wherein the static mixer has an inner diameter of about 1/16 inch to about ½ inch. 26. The system of claim 25, wherein the inner diameter is about ¼ inch. 27. The system of claim 25, wherein the inner diameter is about 1/8 inch. 28. The system of any one of the preceding claims, wherein the polydispersity index of the mixture fluid is about 0.05 to about 0.25. 29. The system of claim 28, wherein the polydispersity index of the mixture fluid is about 0.1 to about 0.2. 30. The system of any one of claims 28, wherein the polydispersity index of the mixture fluid is no greater than about 0.1. 31. The system of any one of the preceding claims, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. 32. The system of claim 31, wherein the Reynolds number is about 500 to about 1000. 33. The system of claim 31, wherein the Reynolds number is about 1000. 34. The system of any one of the preceding claims, wherein the diameter of the lipid nanoparticle is no greater than about 100 nanometers (nm). 35. The system of claim 34, wherein the diameter of the lipid nanoparticle is about 50 nm to about 100 nm. 36. The system of claim 34, wherein the diameter of the lipid nanoparticle is about no greater than about 50 nm. Atty Dkt No.: 58530-742601 37. The system of any one of claims 4-36, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. 38. The system of claim 37, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. 39. The system of claim 37, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. 40. The system of any one of claims 4-39, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. 41. The system of any one of the preceding claims, wherein the static mixer comprises 8 or more elements. 42. The system of any one of the preceding claims, wherein the static mixer comprises 12 or more elements. 43. The system of any one of the preceding claims, wherein the static mixer comprises 16 or more elements. 44. The system of any one of the preceding claims, wherein the static mixer comprises 24 or more elements. 45. The system of any one of the preceding claims, wherein the static mixer is downstream of the conduit. 46. The system of any one of claims 4-45, wherein an angle between the first inlet and the second inlet is no more than 180°. 47. The system of any one of claims 4-46, wherein an angle between the first inlet and the outlet is at least 90°. 48. The system of any one of claims 5-46, wherein an angle between the second inlet and the outlet is at least 90°. 49. The system of any one of the preceding claims, wherein the system reaches a pressure of no greater than 8 pounds per square inch gauge (psig). 50. The system of any one of claims 6-49, further comprising a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate. 51. A method of generating a lipid nanoparticle, the method comprising: (a) providing: (i) a conduit; and Atty Dkt No.: 58530-742601 (ii) a static mixer; (b) providing a nucleic acid and one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate; (c) introducing the nucleic acid to the first inlet; (d) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (e) joining the nucleic acid and the one or more lipid components at the outlet; and (f) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid comprising the lipid nanoparticle. 52. The method of claim 51, wherein the conduit comprises a first inlet. 53. The method of claim 51 or 52, wherein the conduit comprises a second inlet. 54. The method of any one of claims 51-53, wherein the conduit comprises an outlet. 55. A method of generating a lipid nanoparticle, the method comprising: (a) providing: (i) a conduit comprising: (i) a first inlet; (ii) a second inlet; (iii) an outlet; and (ii) a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid, wherein a polydispersity index of the mixture fluid is no greater than about 0.25. 56. A method of generating a lipid nanoparticle, the method comprising: (a) providing: (i) a conduit comprising: (i) a first inlet; (ii) a second inlet; (iii) a junction site downstream of the first inlet and the second inlet; and (ii) a static mixer, wherein the static mixer is disposed at a distal end of the junction site; (b) introducing a nucleic acid to the first inlet; Atty Dkt No.: 58530-742601 (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the junction site; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the nucleic acid and the one or more lipid components do not contact each other prior to entering the junction site. 57. A method of generating a lipid nanoparticle, the method comprising: (a) providing: (i) a conduit comprising: (i) a first inlet; (ii) a second inlet; (iii) an outlet; and (ii) a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby generating a mixture fluid, wherein a Reynolds number of the mixture fluid is no greater than about 1000. 58. A method of generating a lipid nanoparticle, the method comprising: (a) providing: (i) a conduit comprising: (i) a first inlet; (ii) a second inlet; (iii) an outlet; and (ii) a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, wherein a flow rate of the nucleic acid is no greater than about 200 mL/min. 59. A method of generating a lipid nanoparticle, the method comprising: (a) providing: (i) a conduit comprising: Atty Dkt No.: 58530-742601 (i) a first inlet; (ii) a second inlet; (iii) an outlet; and (ii) a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the lipid nanoparticle is no greater than about 150 nm. 60. A method of generating a lipid nanoparticle, the method comprising: (a) providing: (i) a conduit, comprising: (i) a first inlet; (ii) a second inlet; (iii) an outlet; and (ii) a static mixer; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. 61. A method of generating a lipid nanoparticle, the method comprising: (a) providing the system of any one of the preceding claims; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the outlet; and (e) using the static mixer to mix the nucleic acid and the one or more lipid components, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle, wherein a particle size of the lipid nanoparticle is selected by modifying the Reynolds number. Atty Dkt No.: 58530-742601 62. A method of generating a lipid nanoparticle, the method comprising: (a) providing: (i) a conduit comprising: (i) a first inlet; (ii) a second inlet; (iii) a junction site downstream of the first inlet and the second inlet; and (ii) a static mixer, wherein the static mixer is disposed at a distal end of the junction site; (b) introducing a nucleic acid to the first inlet; (c) introducing one or more lipid components of the lipid nanoparticle to the second inlet; (d) joining the nucleic acid and the one or more lipid components at the junction site; and using the static mixer to mix the nucleic acid and the one or more lipid components, wherein the conduit and static mixer reach a pressure of no greater than 8 pounds per square inch gauge (psig). 63. The method of any one of claims 51-62, wherein the conduit comprises a third inlet. 64. The method of any one of claims 51-63, wherein the conduit comprises a fourth inlet. 65. The method of any one of claims 51-64, wherein the conduit is tee-shaped. 66. The method of any one of claims 51-64, wherein the conduit is Y-shaped. 67. The method of any one of claims 51-64, wherein the conduit is L-shaped. 68. The method of any one of claims 51-67, wherein the conduit further comprises a cut zip tie. 69. The method of any one of claims 51-68, wherein the static mixer comprises a plurality of elements, wherein each element of the static mixer makes a 270° twist every 1/8 inch. 70. The method of any one of claims 51-69, wherein the static mixer is less than about 40 mm in length. 71. The method of any one of claims 51-70, wherein the static mixer is less than about 10 mm in length. 72. The method of any one of claims 51-71, wherein the static mixer is less than about 5 mm in length. 73. The method of any one of claims 51-72, wherein the static mixer is about 4.8 mm in length. 74. The method of any one of claims 51-73, wherein the static mixer has an inner diameter of about 1/16 inch to about 1/2 inch. 75. The method of claim 74, wherein the inner diameter is about 1/4 inch. 76. The method of claim 75, wherein the inner diameter is about 1/8 inch. Atty Dkt No.: 58530-742601 77. The method of any one of claims 51-76, wherein the method comprises using a channel housing the static mixer to mix the nucleic acid and the one or more lipid components, wherein the channel is square shaped and makes a -180° twist every 1/8 inch. 78. The method of any one of claims 51-77, wherein the polydispersity index of the mixture fluid is about 0.05 to about 0.25. 79. The method of claim 78, wherein the polydispersity index of the mixture fluid is about 0.1 to about 0.2. 80. The method of claim 78, wherein the polydispersity index of the mixture fluid is no greater than about 0.1. 81. The method of any one of claims 51-80, wherein a Reynolds number of the mixture fluid is about 150 to about 1000. 82. The method of claim 81, wherein the Reynolds number is about 500 to about 1000. 83. The method of claim 82, wherein the Reynolds number is about 1000. 84. The method of any one of claims 81-83, wherein the particle size is increased by decreasing the Reynolds number. 85. The method of any one of claims 81-83, wherein the particle size is decreased by increasing the Reynolds number. 86. The method of any one of claims 81-85, wherein the diameter of the lipid nanoparticle is no greater than about 100 nm. 87. The method of claim 86, wherein the diameter of the lipid nanoparticle is about 50 nm to about 100 nm. 88. The method of claim 86, wherein the diameter of the lipid nanoparticle is about no greater than about 50 nm. 89. The method of any one of claims 51-88, wherein a ratio between a flow rate of the lipid component and a flow rate of the nucleic acid is about 1:2 to about 1:5. 90. The method of claim 89, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:2. 91. The method of claim 89, wherein the ratio between the flow rate of the lipid component and the flow rate of the nucleic acid is about is about 1:3. 92. The method of any one of claims 51-91, wherein the nucleic acid and the one or more lipid components are mixed to generate the mixture fluid, thereby encapsulating the nucleic acid in the one or more lipid components to generate the lipid nanoparticle. Atty Dkt No.: 58530-742601 93. The method of any one of claims 51-92, wherein the static mixer comprises 8 or more elements. 94. The method of any one of claims 51-93, wherein the static mixer comprises 12 or more elements. 95. The method of any one of claims 51-94, wherein the static mixer comprises 16 or more elements. 96. The method of any one of claims 51-95, wherein the static mixer comprises 24 or more elements. 97. The method of any one of claims 51-96, wherein the static mixer is downstream of the conduit. 98. The method of any one of claims 51-97, further comprising diluting the lipid nanoparticle. 99. The method of claim 98, wherein after dilution, a particle size of the lipid nanoparticle is within 10% of the particle size of the lipid nanoparticle before dilution. 100. The method of any one of claims 55-99, further comprising providing the nucleic acid and the one or more lipid components of the lipid nanoparticle to a high performance liquid chromatography (HPLC) pump configured to provide the nucleic acid and the one or more lipid components at a specified flow rate.
PCT/US2025/016069 2024-02-15 2025-02-14 Systems and methods for generating lipid nanoparticles Pending WO2025221360A1 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US202463554054P 2024-02-15 2024-02-15
US63/554,054 2024-02-15
US202463737069P 2024-12-20 2024-12-20
US63/737,069 2024-12-20

Publications (1)

Publication Number Publication Date
WO2025221360A1 true WO2025221360A1 (en) 2025-10-23

Family

ID=97404146

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2025/016069 Pending WO2025221360A1 (en) 2024-02-15 2025-02-14 Systems and methods for generating lipid nanoparticles

Country Status (1)

Country Link
WO (1) WO2025221360A1 (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050196435A1 (en) * 1998-11-13 2005-09-08 Optime Therapeutics, Inc. Method and apparatus for liposome production
US20110204533A1 (en) * 2009-12-22 2011-08-25 Gary Winchester Emulsion-Based Process for Preparing Microparticles and Workhead Assembly for Use with Same
US20190029959A1 (en) * 2015-03-19 2019-01-31 University Of Connecticut Systems and methods for continuous manufacturing of liposomal drug formulations
US20230256436A1 (en) * 2011-10-25 2023-08-17 The University Of British Columbia Limit size lipid nanoparticles and related methods

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050196435A1 (en) * 1998-11-13 2005-09-08 Optime Therapeutics, Inc. Method and apparatus for liposome production
US20110204533A1 (en) * 2009-12-22 2011-08-25 Gary Winchester Emulsion-Based Process for Preparing Microparticles and Workhead Assembly for Use with Same
US20230256436A1 (en) * 2011-10-25 2023-08-17 The University Of British Columbia Limit size lipid nanoparticles and related methods
US20190029959A1 (en) * 2015-03-19 2019-01-31 University Of Connecticut Systems and methods for continuous manufacturing of liposomal drug formulations

Similar Documents

Publication Publication Date Title
Ferhan et al. Lipid nanoparticle technologies for nucleic acid delivery: A nanoarchitectonics perspective
Liu et al. Microfluidic nanoparticles for drug delivery
Okuda et al. On the size-regulation of RNA-loaded lipid nanoparticles synthesized by microfluidic device
Fan et al. Automated high-throughput preparation and characterization of oligonucleotide-loaded lipid nanoparticles
US11745179B2 (en) Microfluidic systems and methods for lipoplex-mediated cell transfection
WO2018190423A1 (en) Flow channel structure and lipid particle or micelle formation method using same
CN223311958U (en) Mixing unit, mixer, microfluidic chip and mixing device
Amrani et al. Characterization of nanoscale loaded liposomes produced by 2D hydrodynamic flow focusing
Debus et al. Optimized preparation of pDNA/poly (ethylene imine) polyplexes using a microfluidic system
Maeki et al. Production of siRNA-loaded lipid nanoparticles using a microfluidic device
WO2025221360A1 (en) Systems and methods for generating lipid nanoparticles
CN119546386A (en) Lipid nanoparticle production system and monitoring and control method thereof
CN114534652A (en) Waveform microstructure mixing unit and application thereof
KR102716110B1 (en) Microfluidic device for preparing scalable lipid nanoparticles, and a process for the preparation thereof
US20130168885A1 (en) Device and method for formation of vesicles
US20240216278A1 (en) Methods for producing nanoparticle dispersions
WO2023186122A1 (en) Countercurrent flow mixing device and method for liposome synthesis
US20230219044A1 (en) Axisymmetric confined impinging jet mixer
EP4596098A1 (en) Microfluidic mixing structure and microfluidic mixing apparatus comprising same
US20240050908A1 (en) Microfluidic platforms for large scale nanoparticle formulations
US20250256248A1 (en) Improved nanocarrier manufacturing
WO2024064072A2 (en) A 3d sheath flow fluidic mixing device for production of nanomaterials
Ahl Microfluidic and Turbulent Mixing for mRNA LNP Vaccines
CN221385957U (en) Inserts and component kits for cross-flow membrane emulsification assemblies
Kim et al. Scalable production of uniform gene-loaded lipid nanoparticles via a fluidity-controlled membrane extrusion