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WO2025215516A1 - Polypeptides glycomodifiés ciblant des auto-anticorps anti-igg4 et leurs utilisations - Google Patents

Polypeptides glycomodifiés ciblant des auto-anticorps anti-igg4 et leurs utilisations

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Publication number
WO2025215516A1
WO2025215516A1 PCT/IB2025/053675 IB2025053675W WO2025215516A1 WO 2025215516 A1 WO2025215516 A1 WO 2025215516A1 IB 2025053675 W IB2025053675 W IB 2025053675W WO 2025215516 A1 WO2025215516 A1 WO 2025215516A1
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WO
WIPO (PCT)
Prior art keywords
sequence
seq
polypeptide
glycoengineered
igg4
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/IB2025/053675
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English (en)
Inventor
Ganesh Venkataraman KAUNDINYA
Tanmoy Chinmoy GANGULY
Melinda Sue HANES
Michela Manni
Jonathan Albert BACK
Brendan REED
Manuela Mally
Kellie COTTER
Tina SCHELBERT
Rainer FOLLADOR
Dominique Nicolas SIRENA
Anna-Janina BEHRENS
Amirreza Faridmoayer
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.)
Glycoera Ag
Original Assignee
Glycoera Ag
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Filing date
Publication date
Application filed by Glycoera Ag filed Critical Glycoera Ag
Publication of WO2025215516A1 publication Critical patent/WO2025215516A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4283—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an allotypic or isotypic determinant on Ig
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
    • C07K2317/41—Glycosylation, sialylation, or fucosylation

Definitions

  • Anti-immunoglobulin G4 (IgG4) autoantibodies are commonly found in autoimmune diseases such as pemphigus vulgaris and membranous nephropathy and are often implicated in disease pathogenesis.
  • the present disclosure identifies certain challenges with existing therapies used to treat diseases associated with immunoglobulin G4 (IgG4) autoantibodies.
  • IgG4 immunoglobulin G4
  • the present disclosure provides a glycoengineered polypeptide comprising: (i) a first moiety comprising one or more peptides that specifically binds to a human immunoglobulin G4 (IgG4 antibody), e.g., as disclosed herein; and (ii) a second moiety comprising one or more glycans conjugated to a first moiety at one or more glycosylation sites, e.g., as disclosed herein.
  • IgG4 antibody human immunoglobulin G4
  • composition comprising a glycoengineered polypeptide as described herein. Further provided herein is a polynucleotide encoding a glycoengineered peptide disclosed herein.
  • This disclosure also provides a Leishmania host cell expressing a glycoengineered polypeptide disclosed herein.
  • a first moiety comprises one or more peptides that binds an IgG4 antibody at a CHI domain, a hinge domain, a CH2 domain, a CH3 domain, or any combination thereof.
  • a first moiety of a glycoengineered polypeptide disclosed herein binds to an IgG4 antibody at a CH2 domain and/or a CH3 domain.
  • a first moiety comprises one or more peptides that binds an IgG4 antibody with a higher affinity as compared to binding to a non-IgG4 antibody.
  • a first moiety binds to an IgG4 autoantibody with an affinity that is at least 10-fold stronger as compared to binding of a first moiety to a non-IgG4 antibody.
  • a non-IgG4 antibody comprises an antibody of a class other than IgG4 (e.g., an IgGl antibody, an IgG2 antibody, an IgG3 antibody, an IgA antibody, an IgM antibody, an IgD antibody, or an IgE antibody, or a fragment or complex of any of the foregoing).
  • a first moiety substantially does not bind to a non-IgG4 antibody.
  • polynucleotides encoding glycoengineered polypeptides, compositions comprising the same, or methods of using the same a first moiety binds to an IgG4 antibody with an affinity of about 1 OpM to about 1 OOOnM.
  • polynucleotides encoding glycoengineered polypeptides, compositions comprising the same, or methods of using the same, a glycoengineered polypeptide is characterized in that administration of a glycoengineered polypeptide to a cell, tissue, or subject reduces a level of an IgG4 antibody as compared to: (1) an otherwise similar cell, tissue, or subject that has not been administered a glycoengineered polypeptide; or (2) the same cell, tissue or subject, prior to administration of a glycoengineered polypeptide.
  • a reduction in IgG4 antibody is at least 5%.
  • a reduction in IgG4 antibody is about 10% to about 99%.
  • polynucleotides encoding glycoengineered polypeptides, compositions comprising the same, or methods of using the same, a glycoengineered polypeptide is characterized in that administration of a glycoengineered polypeptide to a cell, tissue, or subject does not alter (e.g., does not reduce) a level of a non-IgG4 antibody as compared to: (1) an otherwise similar cell, tissue, or subject that has not been administered the glycoengineered polypeptide; or (2) the same cell, tissue or subject, prior to administration of a glycoengineered polypeptide.
  • at least 75% of a non-IgG4 antibody remains (e.g., is detectable) after administration of a glycoengineered polypeptide as compared to a level measured prior to the administration.
  • a first moiety comprises an antibody agent.
  • an antibody agent comprises an antigen binding fragment.
  • an antibody agent comprises a full antibody, a Fab fragment, an scFv, a nanobody, a duobody, or a single domain antibody (e.g., a VHH).
  • an antibody agent of a first moiety comprises one, two or three light chain complementarity determining regions (LC CDRs) and/or one, two or three heavy chain complementary determining regions (HC CDRs).
  • an antibody agent of a first moiety comprises a LC CDR1, a LC CDR2 and a LC CDR3.
  • an antibody agent of a first moiety comprises: (i) a LC CDR1, LC CDR2, and LC CDR3 sequence provided in Table 1; (ii) a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to an LC CDR1, LC CDR2, and LC CDR3 sequence provided in Table 1; or (iii) a sequence having at least 5 substitutions relative to an LC CDR1, LC CDR2, and LC CDR3 sequence provided in Table 1.
  • an antibody agent of a first moiety comprises a HC CDR1, a HC CDR2 and a HC CDR3.
  • an antibody agent of a first moiety comprises: (i) a HC CDR1, HC CDR2, and HC CDR3 sequence provided in Table 2; (ii) a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to an HC CDR1, HC CDR2, and HC CDR3 sequence provided in Table 2; or (iii) a sequence having at least 5 substitutions relative to an HC CDR1, HC CDR2, and HC CDR3 sequence provided in Table 2.
  • an antibody agent of a first moiety comprises (a) a light chain (LC) comprising a sequence provided in Table 1, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity thereto or a sequence having at least 5, 10, or 20 substitutions relative to a VL region provided in Table 1; and/or (b) a heavy chain (HC) comprising a sequence provided in Table 2, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity thereto or a sequence having at least 5, 10, or
  • a first moiety binds to a human IgG4 antibody.
  • a human IgG4 antibody is an IgG4 autoantibody or a fragment or a complex thereof.
  • an IgG4 autoantibody comprises: an anti-PLA2R IgG4 autoantibody or a fragment or a complex thereof; an anti-THSD7A IgG4 autoantibody or a fragment or a complex thereof; an anti-NELL IgG4 autoantibody or a fragment or a complex thereof; an anti -NEP IgG4 autoantibody or a fragment or a complex thereof; an anti -EXT 1 IgG4 autoantibody or a fragment or a complex thereof; and/or an anti-EXT2 IgG4 autoantibody or a fragment or a complex thereof.
  • an IgG4 autoantibody comprises an anti- AD AMTS 13 IgG4 autoantibody or a fragment or a complex thereof.
  • an IgG4 autoantibody comprises an anti-desmoglein 1 IgG4 autoantibody or a fragment or a complex thereof; and/or an anti-desmoglein 3 IgG4 autoantibody or a fragment or a complex thereof.
  • an IgG4 autoantibody comprises an anti-nicotinic acetylcholine receptor (nAChR) IgG4 autoantibody or a fragment or a complex thereof; an anti-muscle- specific kinase (MuSK) IgG4 autoantibody or a fragment or a complex thereof; and/or an anti- low-density lipoprotein receptor-related protein 4 (LRP4) IgG4 autoantibody or a fragment or a complex thereof.
  • nAChR non-nicotinic acetylcholine receptor
  • MoSK anti-muscle- specific kinase
  • LRP4 anti- low-density lipoprotein receptor-related protein 4
  • a second moiety comprises one or more glycans that specifically bind to one or more endocytic receptors.
  • an endocytic receptor is or comprises an endocytic lectin.
  • an endocytic receptor is chosen from: an asialoglycoprotein receptor (ASGPR); a mannose binding receptor, a Cluster of Differentiation 206 (CD206) receptor; a DC-SIGN (Cluster of Differentiation 209 or CD209) receptor; a C-Type Lectin Domain Family 4 Member G (LSECTin) receptor; a macrophage inducible Ca2+- dependent lectin receptor (Mincle); a L-SIGN CD209L receptor; dectin- 1; dectin -2, langerin, macrophage mannose 2 receptor, BDCA-2, DCIR, MBL, MDL, MICL, CLEC2, DNGR1, CLEC12B, DEC-205, and mannose 6 phosphate receptor (M6PR), or a combination thereof.
  • ASGPR asialoglycoprotein receptor
  • CD206 Cluster of Differentiation 206
  • DC-SIGN Cluster of Differentiation 209 or CD209
  • LSECTin C-Type Lect
  • a second moiety comprises a glycan structure comprising a biantennary GalNAc.
  • a biantennary GalNac binds to an asialoglycoprotein receptor (ASGPR) or a fragment or variant thereof, or a complex comprising ASGPR.
  • ASGPR asialoglycoprotein receptor
  • an N-glycan has a structure of wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N-acetylglucosamine (GlcNAc) residue and the black circle represents a mannose (Man) residue, and wherein X represents an amino acid residue of a first moiety.
  • an N-glycan is conjugated to a glycoengineered polypeptide at least one, two, three, or four N-glycosylation sites. In some embodiments, an N-glycan is conjugated to a glycoengineered polypeptide at one, two, three, or four N-glycosylation sites. In some embodiments, an N-glycosylation site comprises a consensus sequence of N-X-S/T or N-X-C, wherein X is any amino acid except proline. In some embodiments, an N-glycosylation site is naturally occurring.
  • an N-glycosylation site is engineered into an amino acid sequence of a first moiety.
  • an N-glycosylation site comprises a sequence of SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 168, or SEQ ID NO: 169.
  • an endocytic receptor is or comprises ASGPR or a fragment or variant thereof, or a complex comprising ASGPR.
  • a glycan structure of a second moiety comprises a terminal GalNac.
  • a glycoengineered polypeptide is characterized in that when administered to a cell, tissue, or subject, a glycoengineered polypeptide which is bound to an IgG4 antibody (e.g., IgG4 autoantibody) or a complex comprising same via a first moiety, and to an endocytic receptor via a second moiety results in degradation of a IgG4 antibody or a complex comprising same.
  • IgG4 antibody e.g., IgG4 autoantibody
  • degradation comprises internalization into a cell.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • an immune complex comprising same and a glycoengineered polypeptide are internalized into a cell.
  • internalization occurs in an ASGPR-dependent manner.
  • a magnitude of internalization depends on an expression and/or an activity level of ASGPR.
  • degradation comprises lysosomal degradation.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • an immune complex comprising same, and a glycoengineered polypeptide are internalized into a lysosome.
  • a composition disclosed herein comprises a population of glycoengineered polypeptides as disclosed herein.
  • a population of glycoengineered polypeptides has an N-glycan profile that is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or about 100% homogeneous at one or more N-glycosylation site(s).
  • composition disclosed herein is a pharmaceutical composition.
  • the present disclosure further provides a method comprising delivering a pharmaceutical composition as disclosed herein to a cell, tissue, or subject.
  • IgG4 antibody e.g., an IgG4 autoantibody
  • a complex comprising same comprising delivering a pharmaceutical composition as disclosed herein to a cell, tissue, or subject.
  • Also provided herein is a method of delivering an IgG4 antibody (e.g., an IgG4 autoantibody) or a complex comprising same to a lysosome comprising delivering a pharmaceutical composition as disclosed herein to a cell, tissue, or subject.
  • an IgG4 antibody e.g., an IgG4 autoantibody
  • a complex comprising same to a lysosome comprising delivering a pharmaceutical composition as disclosed herein to a cell, tissue, or subject.
  • any of the methods disclosed herein further comprise administering a pharmaceutical composition to a cell, tissue, or subject.
  • a subject has or is diagnosed as having a disease associated with one or more IgG4 autoantibodies.
  • a disease associated with one or more IgG4 autoantibodies comprises: Pemphigus Vulgaris/Folaceus; Muscle-specific kinase (MuSK) myasthenia gravis (MG), Idiopathic Thrombotic Thrombocytopenia Purpura (iTTP), idiopathic membranous nephropathy (iMN), Chronic Inflammatory Demyelinating Polyneuropathy (CIDP), or Immunoglobulin G4- Related Disease (IgG4-RD).
  • Muscle-specific kinase MoSK
  • MG myasthenia gravis
  • iTTP Idiopathic Thrombotic Thrombocytopenia Purpura
  • iMN idiopathic membranous nephropathy
  • CIDP Chronic Inflammatory Demyelinating Polyneuropathy
  • IgG4-RD Immunoglobulin G4- Related Disease
  • administration of a glycoengineered polypeptide disclosed herein or a pharmaceutical compositions comprising the same to a subject reduces a level of an IgG4 autoantibody as compared to a subject who has not been administered a pharmaceutical composition or as compared to the same subject prior to administration of a pharmaceutical composition.
  • a reduction in IgG4 antibody is at least 5%. In some embodiments, a reduction in IgG4 antibody is about 10% to about 99%.
  • administration of a glycoengineered polypeptide disclosed herein or a pharmaceutical compositions comprising the same to a subject substantially does not reduce a level of a non-IgG4 immunoglobulin as compared to a subject who has not been administered a pharmaceutical composition or as compared to the same subject prior to administration of a pharmaceutical composition.
  • at least 75% of a non-IgG4 immunoglobulin remains (e.g., is detectable) after administration of a pharmaceutical composition.
  • administration of a pharmaceutical composition reduces less than 10% of a non- IgG4 immunoglobulin as compared to a subject who has not been administered a pharmaceutical composition or as compared to the same subject prior to administration of a pharmaceutical composition.
  • a non-IgG4 immunoglobulin is an immunoglobulin of a different IgG class as compared to IgG4.
  • a non-IgG4 immunoglobulin is an immunoglobulin of a different isotype as compared to IgG4.
  • a non-IgG4 immunoglobulin is or comprises IgGl, IgG2, IgG3, IgA, IgM, IgD or IgE, or a fragment or complex of any of the foregoing.
  • glycoengineered polypeptides disclosed herein nucleic acids encoding the same, composition comprising glycoengineered polypeptides or nucleic acids encoding the same, and methods of making and using the same are provided throughout the present disclosure.
  • FIGS. 1A-1C show immunodepletion of IgG isotypes from pooled healthy donor serum.
  • Exemplary anti-IgG4 antibody agents were coated on magnetic beads followed by incubation with healthy donor serum. Samples were next placed on a magnet and supernatant collected to obtain the depleted sera. IgG content was measured by multiplex ELISA. Representative depletion of IgG isotypes is shown for a representative anti-IgG4 antibody agent from Bin 3 (FIG. 1A) and two distinct representative anti-IgG4 antibody agents from Bin-1 (FIGS. 1B-1C).
  • FIGS. 2A-2B show immunodepletion of autoantibodies from pMN serum.
  • Exemplary anti-IgG4 antibody agents were coated on beads followed by incubation with pMN serum. Samples were next placed on a magnet and supernatant collected to obtain the depleted sera.
  • Anti-PLA2R autoantibody in sample was evaluated by ELISA. Representative depletion of autoantibodies from the sera of 3 pMN patients for two distinct anti-IgG4 antibody agents in bin- 1 is shown in FIGS. 2A and 2B.
  • FIGS. 3A-3C show exemplary glycoengineered polypeptide binding to IgG4.
  • Exemplary glycoengineered polypeptides binding to human purified IgG4 was evaluated by ELISA. Representative binding to human purified IgG4 are shown for a representative for each bin: FIG. 3A, bin-1; FIG. 3B, bin-2; and FIG. 3C, bin-3.
  • An exemplary glycoengineered polypeptide is also referred to as a “G-LyTAC” herein.
  • FIG. 4 is a graph depicting binding affinity from a Surface Plasmon Resonance (SPR) assay for an exemplary glycoengineered anti-IgG4 Fab polypeptide in bin-3 for human IgG4 (hIgG4).
  • SPR Surface Plasmon Resonance
  • FIG. 5 is a panel of graphs depicting binding of a representative anti-IgG4 antibody agent from bin 3 and a corresponding exemplary glycoengineered polypeptide to different IgG subclasses and Ig isotypes.
  • FIG. 6 shows binding kinetics of an exemplary glycoengineered anti-IgG4 Fab polypeptide from bin-3 to ASGPR1 as determined by SPR.
  • FIGS. 7A-7B depict glycan content and purity analysis of exemplary glycoengineered polypeptides.
  • FIGS. 8A-8B show the internalization of an exemplary anti-IgG4 glycoengineered polypeptides by fluorescence microscopy.
  • FIG. 8A is a panel of microscopy images.
  • FIG. 8B is a graph showing internalization of the exemplary glycoengineered anti-IgG4 polypeptide across multiple doses (as indicated) and over time.
  • data points with 500nM are displayed with the top most line, followed by data points with 166.6nM and data points with 55.5nM (as indicated in the graph).
  • FIGS. 9A-9C depict internalization of exemplary glycoengineered anti-IgG4 polypeptides in HepG2 cells.
  • Exemplary glycoengineered anti-IgG4 polypeptides were labelled with a pH sensitive dye (pHrodo) to monitor internalization and localization in the lysosome.
  • HepG2 wt, ASGPRlko and ASGPR2ko cells were incubated with pHrodo-labelled- exemplary glycoengineered polypeptides for 3 hours followed by evaluation of internalization by flow cytometry. Representative data for 3 distinct binders from bin-2 are shown in FIGS. 9A, 9B, and 9C.
  • FIGS. 10A-10C show degradation of IgG4 in HepG2 cells.
  • HepG2 cells were incubated with exemplary glycoengineered anti-IgG4 polypeptides alone or in complex with human purified IgG4 or recombinant IgG4 for 4h (TO). HepG2 cells were then washed and incubated for an additional l-24h to allow for complex degradation. Internalization of IgG4 was monitored by Western Blot. Representative data is shown for a bin-3 exemplary glycoengineered anti-IgG4 polypeptide (FIG. 10A), and two distinct exemplary glycoengineered anti-IgG4 polypeptides from Bin 1 (FIGS. 10B-C).
  • FIG. 11 is a graph showing IgG4 polypeptide levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin 3.
  • FIG. 12 is a graph showing IgG4 polypeptide levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin 1.
  • FIG. 13 is a graph showing IgG4 polypeptide levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin 2.
  • FIGS. 14A-14F are graphs showing in vivo selective depletion of IgG4 in animals administered exemplary glycoengineered anti-IgG4 polypeptides.
  • FIG. 14A IgG4 levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin-3.
  • FIG. 14B IgGl levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin-3.
  • FIG. 14C IgGl levels in animals administered an exemplary glycoengineered anti- IgG4 polypeptide from bin-1.
  • FIG. 14D IgG2 levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin-1.
  • FIG. 14E IgG3 levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin-1.
  • FIG. 14F IgG4 levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin-1.
  • FIGS. 15A-15B are each a pair of graphs depicting the levels of liver enzymes in animals administered an exemplary glycoengineered anti-IgG4 polypeptide.
  • FIG. 15A alanine aminotransferase (ALT) (left) and alkaline phosphatase (AP)(right) levels in animals administered (i.v.) different doses of an exemplary glycoengineered polypeptide from bin-2 or PBS vehicle.
  • FIG. 15B Aspartate aminotransferase (ASP) (left) and gamma-glutamyl transferase (GGT) (right) levels in animals administered (i.v.) different doses of an exemplary glycoengineered anti-IgG4 polypeptide from bin-2 or PBS vehicle.
  • ASP Aspartate aminotransferase
  • GTT gamma-glutamyl transferase
  • FIG. 16 is a set of graphs depicting blood chemistry in animals administered an exemplary glycoengineered anti-IgG4 polypeptide.
  • FIG. 16 depicts levels of creatine kinase (left panel), albumin (middle panel) and bilirubin (right panel), in animals administered an exemplary glycoengineered polypeptide from bin-2 (at an i.v. dose of 5mg/kg, 12.5 mg/kg, or 25mg/kg), or PBS vehicle.
  • FIGS. 17A-17B are graphs depicting depletion of IgG4 in animals administered an exemplary glycoengineered anti-IgG4 polypeptide.
  • FIG. 17A IgG4 levels in animals administered an exemplary glycoengineered anti-IgG4 polypeptide from bin-1 intravenously at different doses compared to animals administered PBS only. Serum samples were collected at the timepoints indicated and total IgG4 levels were quantified by ELISA. The graph shows the average ⁇ SEM of % of IgG4 target remaining in serum, normalized to predose level, from
  • FIGS. 18A-18B show tissue distribution of an exemplary glycoengineered anti-IgG4 polypeptide.
  • FIG. 18A Representative images of tissue distribution of an exemplary glycoengineered anti-IgG4 polypeptide from bin-1 assessed by ex vivo imaging of tissues collected at 15 min, 1 hour, and 72 hours.
  • FIG. 18B Quantification of tissue distribution of the exemplary glycoengineered anti-IgG4 polypeptide from bin-1 at 1 hour.
  • the y-axis represents fluorescence (Ph/s/cm 2 /sr).
  • FIG. 19 shows a graph depicting the in vivo depletion of pathogenic pMN patient- derived autoantibodies using an exemplary glycoengineered anti-IgG4 polypeptide from bin-1.
  • the term “a” may be understood to mean “at least one”; (ii) the term “or” may be understood to mean “and/or”; (iii) the terms “comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps; and (iv) the terms “about” and “approximately” may be understood to permit standard variation as would be understood by those of ordinary skill in the art; and (v) where ranges are provided, endpoints are included.
  • Glycans refers to one or more saccharides or sugar chains that can be attached to a protein or lipid to form a glycoconjugate.
  • a glycan conjugated to a protein forms a glycoprotein.
  • a glycan conjugated to a nitrogen atom of an amino acid residue is an N-linked glycan and a glycan conjugated to an oxygen atom of an amino acid residue is an O-linked glycan.
  • the structure of a glycan indicates if a specific glycan is an N-linked glycan.
  • Glycoengineered means a process of glycosylating a target protein (e.g., a glycoengineered polypeptide disclosed herein), or a target protein made by such process.
  • the process uses a host cell system that has one or more enzymes (e.g., pathways) that provides for glycosylation of the target protein; in some other embodiments, the process is performed by chemically attaching one or more glycans to a target protein, e.g., using Click chemistry.
  • a host cell system can be genetically engineered to introduce a glycosylation pathway to selectively glycosylate a target protein with a particular glycan structure.
  • a host cell used to generate a glycoengineered target protein can include, for example, a recombinant nucleic acid encoding a target protein; and a recombinant nucleic acid encoding a heterologous glycosyltransferase.
  • the host cell system used for glycoengineering e.g., to generate a glycoengineered protein
  • the host cell system used for glycoengineering e.g, to generate a glycoengineered protein
  • the host cell used for glycoengineering or to generate a glycoengineered target protein can be a mammalian cell, an insect cell, a yeast cell, a bacterial cell, a plant cell, a microalgae, or a protozoa.
  • the protozoa used for glycoengineering can be a species of Leishmania.
  • a glycoengineered target protein also includes a target protein that has been engineered to be selectively glycosylated at one or more specific sites when generated in the host cell system.
  • Glycoengineered polypeptide As used herein, a “glycoengineered polypeptide” is a polypeptide that mediates the internalization and/or degradation of a target protein by specifically binding to a target protein (e.g., an IgG4 antibody) and engaging with one or more endocytic receptors. In some embodiments, binding (e.g., simultaneous binding) of a glycoengineered polypeptide to a target protein and an endocytic receptor internalizes a target protein and/or activates one or more degradation pathways.
  • a target protein e.g., an IgG4 antibody
  • Glycosylation site refers to a site of glycosylation in a protein. Such a glycosylation site, also referred to as a glycosite herein, can be naturally present in the amino acid sequence of a protein or recombinantly engineered into the protein by addition or substitution or deletion of amino acids. In some embodiments, a glycosylation site is present in a so-called glycotag that is fused to a glycoengineered polypeptide disclosed herein. In certain embodiments, a glycotag is fused to a protein to create a bispecific binding protein.
  • a glycotag refers to a peptide containing consensus N- glycosylation site sequence fused to N- or a C-terminal or both termini of a protein or polypeptide.
  • the glycotag is fused to the C-terminus of the of the glycoengineered polypeptide disclosed herein via a peptide linker.
  • the glycotag is fused to the N-terminus of the glycoengineered polypeptide disclosed herein via a peptide linker.
  • the peptide linker is a consensus peptide sequence.
  • the consensus peptide sequence is 1, 2, 3, 4, 5, 6, 7 or more amino acid residues in length.
  • the bifunctional protein provided herein contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more glycotags.
  • Endocytic receptor refers to a receptor or a fragment thereof that binds to a target and internalizes the target into a cell.
  • an endocytic receptor recognizes and binds to one or more glycans on a target.
  • binding of an endocytic receptor to a target internalizes the target into a cell, e.g., into a lysosome or phagosome.
  • an endocytic receptor is or comprises an endocytic lectin.
  • an endocytic receptor is chosen from: an asialoglycoprotein receptor (ASGPR); a mannose binding receptor, a Cluster of Differentiation 206 (CD206) receptor; a DC-SIGN (Cluster of Differentiation 209 or CD209) receptor; a C-Type Lectin Domain Family 4 Member G (LSECTin) receptor; a macrophage inducible Ca2+- dependent lectin receptor (Mincle); a L-SIGN CD209L receptor; dectin- 1; dectin -2, langerin, macrophage mannose 2 receptor, BDCA-2, DCIR, MBL, MDL, MICL, CLEC2, CLEC10, DNGR1, CLEC12B, DEC-205, and mannose 6 phosphate receptor (M6PR), or a combination thereof.
  • ASGPR asialoglycoprotein receptor
  • CD206 Cluster of Differentiation 206
  • DC-SIGN Cluster of Differentiation 209 or CD209
  • LSECTin C-
  • Administration typically refers to the administration of a composition to a subject or system, for example to achieve delivery of an agent that is, or is included in or otherwise delivered by, the composition.
  • an animal is a domestic animal, such as a companion animal, e.g., a dog or a cat; in some embodiments, an animal is an animal used in agriculture (e.g., farming [e.g., a cow, a sheep or a horse]) or for recreation.
  • administration may be systemic or local.
  • bronchial e.g., by bronchial instillation
  • buccal dermal
  • dermal which may be or comprise, for example, one or more of topical to the dermis, intradermal, interdermal, transdermal, etc
  • enteral intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a specific organ (e. g.
  • administration may be by injection (e.g., intramuscular, intravenous, or subcutaneous injection).
  • injection may involve bolus injection, drip, perfusion, or infusion.
  • administration may involve only a single dose.
  • administration may involve application of a fixed number of doses.
  • administration may involve dosing that is intermittent (e.g., a plurality of doses separated in time) and/or periodic (e.g., individual doses separated by a common period of time) dosing. In some embodiments, administration may involve continuous dosing (e.g., perfusion) for at least a selected period of time.
  • an antibody agent can be formulated for oral delivery. For example, one with skill in the art will understand that an antibody agent disclosed herein can be formulated for oral delivery using technologies developed by Oramed (https://www.oramed.com/) or Premas (https://www.premasbiotech.com/).
  • adult refers to a human eighteen years of age or older. In some embodiments, a human adult has a weight within the range of about 90 pounds to about 250 pounds.
  • a first moiety comprising one or more peptides that specifically bind to a target autoantibody (e.g., an IgG4 autoantibody), has a high affinity to an autoantigen (e.g., a IgG4 autoantigen).
  • a target autoantibody e.g., an IgG4 autoantibody
  • an autoantigen e.g., a IgG4 autoantigen
  • a high affinity is an affinity of about 50pM to about 200pM.
  • affinity is assessed in a quantitative assay.
  • a moderate affinity is an affinity of about 0.201nM to about 6nM.
  • affinity is assessed in a quantitative assay.
  • affinity is assessed over a plurality of concentrations (e.g., of one binding partner at a time).
  • affinity is assessed in the presence of one or more potential competitor entities (e.g., that might be present in a relevant - e.g., physiological - setting).
  • affinity is assessed relative to a reference (e.g., that has a known affinity above a particular threshold [a “positive control” reference] or that has a known affinity below a particular threshold [ a “negative control” reference”].
  • affinity may be assessed relative to a contemporaneous reference; in some embodiments, affinity may be assessed relative to a historical reference. Typically, when affinity is assessed relative to a reference, it is assessed under comparable conditions.
  • Avidity As is known in the art, “avidity” is a measure of the accumulated strength of multiple non-covalent interactions between two or more binding partners in a complex.
  • avidity can be determined by (1) a binding affinity of two or more binding partners in a complex; (2) valency of each of the binding partners in a complex; and/or (3) structural arrangements of two or more binding partners in a complex.
  • the avidity of binding between two or more binding partners is more than a sum of each binding affinity between the two or more binding partners.
  • avidity is also referred to as apparent affinity or functional affinity.
  • a second moiety comprising one or more glycans which can bind to a receptor (e.g., an endocytic receptor) contributes to binding avidity of the glycoengineered polypeptide.
  • an endocytic receptor is ASGPR or a fragment or variant thereof.
  • avidity is assessed in a quantitative assay. In some embodiments, avidity is assessed over a plurality of concentrations. In some embodiments, avidity is assessed in the presence of one or more potential competitor entities (e.g., that might be present in a relevant - e.g., physiological - setting). In some embodiments, avidity may be assessed relative to a contemporaneous reference; in some embodiments, avidity may be assessed relative to a historical reference. Typically, when avidity is assessed relative to a reference, it is assessed under comparable conditions.
  • agent may refer to a physical entity or phenomenon. In some embodiments, an agent may be characterized by a particular feature and/or effect. In some embodiments, an agent may be a compound, molecule, or entity of any chemical class including, for example, a small molecule, polypeptide, nucleic acid, saccharide, lipid, metal, or a combination or complex thereof. In some embodiments, the term “agent” may refer to a compound, molecule, or entity that comprises a polymer. In some embodiments, the term may refer to a compound or entity that comprises one or more polymeric moieties.
  • the term “agent” may refer to a compound, molecule, or entity that is substantially free of a particular polymer or polymeric moiety. In some embodiments, the term may refer to a compound, molecule, or entity that lacks or is substantially free of any polymer or polymeric moiety.
  • Amino acid in its broadest sense, as used herein, refers to any compound and/or substance that can be incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds.
  • an amino acid has the general structure H2N- C(H)(R)-COOH. In some embodiments, an amino acid is a naturally- occurring amino acid.
  • an amino acid is a non-natural amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L-amino acid.
  • Standard amino acid refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides.
  • Nonstandard amino acid refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source.
  • an amino acid, including a carboxy- and/or amino-terminal amino acid in a polypeptide can contain a structural modification as compared with the general structure above.
  • an amino acid may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and/or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and/or the hydroxyl group) as compared with the general structure.
  • such modification may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino acid.
  • such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino acid, as compared with one containing an otherwise identical unmodified amino acid.
  • the term “amino acid” may be used to refer to a free amino acid; in some embodiments it may be used to refer to an amino acid residue of a polypeptide.
  • Animal refers to a member of the animal kingdom.
  • “animal” refers to humans; unless otherwise specified, in many embodiments, a human may be of either gender and/or at any stage of development.
  • “animal” refers to non-human animals; unless otherwise specified, in many embodiments, a nonhuman animal may be of any gender and/or at any stage of development.
  • a non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, and/or a pig).
  • an animal may be, for example, a mammal, a bird, a reptile, an amphibian, a fish, an insect, a worm, etc..
  • an animal may be a transgenic animal, genetically engineered animal, and/or a clone.
  • Antibody agent refers to a polypeptide that includes canonical immunoglobulin sequence elements sufficient to confer specific binding to a particular target antigen.
  • the antigen that is bound by the anti-IgG4 antibody agent is an IgG4 antibody (e.g., an IgG4 autoantibody).
  • IgG4 antibody e.g., an IgG4 autoantibody.
  • intact antibodies as produced in nature are approximately 150 kD tetrameric agents comprised of two identical heavy chain polypeptides (about 50 kD each) and two identical light chain polypeptides (about 25 kD each) that associate with each other into what is commonly referred to as a “Y-shaped” structure.
  • Each heavy chain is comprised of at least four domains (each about 110 amino acids long)- an amino-terminal variable (VH) domain (located at the tips of the Y structure), followed by three constant domains: CHI, CH2, and the carboxy -terminal CH3 (located at the base of the Y’s stem).
  • VH amino-terminal variable
  • CHI amino-terminal variable
  • CH2 amino-terminal variable
  • CH3 located at the base of the Y’s stem
  • a short region known as the “switch” connects the heavy chain variable and constant regions.
  • the “hinge” connects CH2 and CH3 domains to the rest of the antibody. Two disulfide bonds in this hinge region connect the two heavy chain polypeptides to one another in an intact antibody.
  • Each light chain is comprised of two domains - an amino-terminal variable (VL) domain, followed by a carboxy-terminal constant (CL) domain, separated from one another by another “switch”.
  • Intact antibody tetramers are comprised of two heavy chain-light chain dimers in which the heavy and light chains are linked to one another by a single disulfide bond; two other disulfide bonds connect the heavy chain hinge regions to one another, so that the dimers are connected to one another and the tetramer is formed.
  • Naturally-produced antibodies are also glycosylated, typically on the CH2 domain.
  • Each domain in a natural antibody has a structure characterized by an “immunoglobulin fold” formed from two beta sheets (e.g., 3-, 4-, or 5-stranded sheets) packed against each other in a compressed antiparallel beta barrel.
  • Each variable domain contains three hypervariable loops known as “complementarity determining regions” (CDR1, CDR2, and CDR3) and four somewhat invariant “framework” regions (FR1, FR2, FR3, and FR4).
  • CDR1, CDR2, and CDR3 three hypervariable loops known as “complementarity determining regions” (CDR1, CDR2, and CDR3) and four somewhat invariant “framework” regions (FR1, FR2, FR3, and FR4).
  • the Fc region of naturally-occurring antibodies binds to elements of the complement system, and also to receptors on effector cells, including for example effector cells that mediate cytotoxicity.
  • affinity and/or other binding attributes of Fc regions for Fc receptors can be modulated through glycosylation or other modification.
  • antibodies produced and/or utilized in accordance with the present disclosure include glycosylated Fc domains, including Fc domains with modified or engineered such glycosylation.
  • antibodies produced and/or utilized in accordance with the present disclosure include one or more modifications on an Fc domain, e.g., an effector null mutation, e.g., a LALA, LAGA, FEGG, AAGG, or AAGA mutation.
  • an effector null mutation e.g., a LALA, LAGA, FEGG, AAGG, or AAGA mutation.
  • any polypeptide or complex of polypeptides that includes sufficient immunoglobulin domain sequences as found in natural antibodies can be referred to and/or used as an “antibody”, whether such polypeptide is naturally produced (e.g., generated by an organism reacting to an antigen), or produced by recombinant engineering, chemical synthesis, or other artificial system or methodology.
  • an antibody is polyclonal; in some embodiments, an antibody is monoclonal. In some embodiments, an antibody has constant region sequences that are characteristic of dog, cat, mouse, rabbit, primate, or human antibodies. In some embodiments, antibody sequence elements are human, humanized, primatized, chimeric, etc, as is known in the art. Moreover, the term “antibody” as used herein, can refer in appropriate embodiments (unless otherwise stated or clear from context) to any of the art-known or developed constructs or formats for utilizing antibody structural and functional features in alternative presentation.
  • an antibody utilized in accordance with the present invention is in a format selected from, but not limited to, intact IgA, IgG, IgE or IgM antibodies; bi- or multi- specific antibodies (e.g., Zybodies®, etc); antibody fragments such as Fab fragments, Fab’ fragments, F(ab’)2 fragments, Fd’ fragments, Fd fragments, and isolated CDRs or sets thereof; single chain Fvs; polypeptide-Fc fusions; single domain antibodies (e.g., VHH [e.g., a camelid VHH] or NAR) alternative scaffolds or antibody mimetics (e.g., anticalins, FN3 monobodies, DARPins, Affibodies, Affilins, Affimers, Affitins, Alphabodies, Avimers, Fynomers, Im7, VLR, VNAR, Trimab, CrossMab, Trident); nanobodies,
  • relevant formats may be or include: Adnectins®; Affibodies®; Affilins®; Anticalins®; Avimers®; BiTE®s; cameloid antibodies; Centyrins®; ankyrin repeat proteins or DARPINs®; dual-affinity re-targeting (DART) agents; Fynomers®; shark single domain antibodies such as IgNAR; immune mobilixing monoclonal T cell receptors against cancer (ImmTACs); KALBITOR®s; MicroProteins; Nanobodies® minibodies; masked antibodies (e.g., Probodies®); Small Modular ImmunoPharmaceuticals (“SMIPsTM ); single chain or Tandem diabodies (TandAb®); TCR-like antibodies;, Trans-bodies®; TrimerX®; VHHs.
  • Adnectins® Adnectins®
  • Affibodies® Affilins®
  • Anticalins® Anticalins®
  • Avimers®
  • an antibody may lack a covalent modification (e.g., attachment of a glycan) that it would have if produced naturally.
  • an antibody format is or comprises a VHH, e.g., a camelid VHH.
  • a VHH is a multivalent VHH, e.g., a bivalent VHH.
  • an antibody comprises a single domain antibody, e.g., comprising one or more additional domains such as an Fc, a half-Fc (e.g., comprising an interchain cysteine mutant), an albumin domain, or combinations thereof.
  • an antibody comprises a single chain Fv, e.g., comprising one or more additional domains such as an Fc, a half-Fc (e.g., comprising an interchain cysteine mutant), an albumin domain, or combinations thereof.
  • an antibody comprises a polypeptide- Fc fusion.
  • an antibody may contain a covalent modification (e.g., attachment of a glycan, a pay load [e.g., a detectable moiety, a therapeutic moiety, a catalytic moiety, etc], or other pendant group [e.g., poly-ethylene glycol, etc.]).
  • an “antibody fragment” refers to a portion of an antibody or antibody agent as described herein and typically refers to a portion that includes an antigen-binding portion or variable region thereof.
  • An antibody fragment may be produced by any means. For example, in some embodiments, an antibody fragment may be enzymatically or chemically produced by fragmentation of an intact antibody or antibody agent. Alternatively, in some embodiments, an antibody fragment may be recombinantly produced (e.g., by expression of an engineered nucleic acid sequence. In some embodiments, an antibody fragment may be wholly or partially synthetically produced.
  • an antibody fragment (particularly an antigen-binding antibody fragment) may have a length of at least about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 amino acids or more, in some embodiments at least about 200 amino acids.
  • Antigen refers to an agent that elicits an immune response; and/or (ii) an agent that binds to a T cell receptor (e.g., when presented by an MHC molecule) or to an antibody.
  • an antigen elicits a humoral response (e.g., including production of antigen-specific antibodies); in some embodiments, an elicits a cellular response (e.g., involving T-cells whose receptors specifically interact with the antigen).
  • and antigen binds to an antibody and may or may not induce a particular physiological response in an organism.
  • an antigen may be or include any chemical entity such as, for example, a small molecule, a nucleic acid, a polypeptide, a carbohydrate, a lipid, a polymer (in some embodiments other than a biologic polymer [e.g., other than a nucleic acid or amino acid polymer) etc.
  • an antigen is or comprises a polypeptide.
  • an antigen is or comprises a glycan.
  • an antigen may be provided in isolated or pure form, or alternatively may be provided in crude form (e.g., together with other materials, for example in an extract such as a cellular extract or other relatively crude preparation of an antigen-containing source).
  • antigens utilized in accordance with the present invention are provided in a crude form.
  • an antigen is a recombinant antigen.
  • Binding typically refers to a non-covalent association between or among two or more entities. “Direct” binding involves physical contact between entities or moieties; indirect binding involves physical interaction by way of physical contact with one or more intermediate entities. Binding between two or more entities can typically be assessed in any of a variety of contexts - including where interacting entities or moieties are studied in isolation or in the context of more complex systems (e.g., while covalently or otherwise associated with a carrier entity and/or in a biological system or cell).
  • CDR refers to a complementarity determining region within an antibody variable region. There are three CDRs in each of the variable regions of the heavy chain and the light chain, which are designated CDR1, CDR2 and CDR3, for each of the variable regions.
  • a "set of CDRs” or “CDR set” refers to a group of three or six CDRs that occur in either a single variable region capable of binding the antigen or the CDRs of cognate heavy and light chain variable regions capable of binding the antigen.
  • composition may be used to refer to a discrete physical entity that comprises one or more specified components.
  • a composition may be of any form - e.g., gas, gel, liquid, solid, etc.
  • composition or method described herein as “comprising” one or more named elements or steps is open-ended, meaning that the named elements or steps are essential, but other elements or steps may be added within the scope of the composition or method.
  • any composition or method described as “comprising” (or which "comprises") one or more named elements or steps also describes the corresponding, more limited composition or method “consisting essentially of' (or which "consists essentially of') the same named elements or steps, meaning that the composition or method includes the named essential elements or steps and may also include additional elements or steps that do not materially affect the basic and novel characteristic(s) of the composition or method.
  • composition or method described herein as “comprising” or “consisting essentially of' one or more named elements or steps also describes the corresponding, more limited, and closed-ended composition or method "consisting of' (or “consists of') the named elements or steps to the exclusion of any other unnamed element or step.
  • known or disclosed equivalents of any named essential element or step may be substituted for that element or step.
  • Conjugate refers to linking of one moiety to another moiety by in vitro methods (e.g., chemical synthesis) or in vivo (e.g., in a cell).
  • a moiety comprising one or more glycans e.g., a second moiety
  • a moiety comprising one or more glycans is conjugated to a different moiety, for example, at one or more glycosylation sites in vivo in a cell.
  • a moiety comprising one or more glycans is conjugated to a different moiety, for example, at one or more glycosylation sites by chemical conjugation.
  • Domain refers to a section or portion of an entity.
  • a “domain” is associated with a particular structural and/or functional feature of the entity so that, when the domain is physically separated from the rest of its parent entity, it substantially or entirely retains the particular structural and/or functional feature.
  • a domain may be or include a portion of an entity that, when separated from that (parent) entity and linked with a different (recipient) entity, substantially retains and/or imparts on the recipient entity one or more structural and/or functional features that characterized it in the parent entity.
  • a domain is a section or portion of a molecule (e.g., a small molecule, carbohydrate, lipid, nucleic acid, or polypeptide).
  • a domain is a section of a polypeptide; in some such embodiments, a domain is characterized by a particular structural element (e.g., a particular amino acid sequence or sequence motif, alpha-helix character, alpha-sheet character, coiled-coil character, random coil character, etc.), and/or by a particular functional feature (e.g., binding activity, enzymatic activity, folding activity, signaling activity, etc.).
  • Epitope includes any moiety that is specifically recognized by an immunoglobulin (e.g., antibody or receptor) binding component.
  • an epitope is comprised of a plurality of chemical atoms or groups on an antigen.
  • such chemical atoms or groups are surface-exposed when the antigen adopts a relevant three-dimensional conformation.
  • such chemical atoms or groups are physically near to each other in space when the antigen adopts such a conformation.
  • at least some such chemical atoms are groups are physically separated from one another when the antigen adopts an alternative conformation (e.g., is linearized).
  • a “functional” biological molecule is a biological molecule in a form in which it exhibits a property and/or activity by which it is characterized.
  • Fragment A “fragment” of a material or entity as described herein has a structure that includes a discrete portion of the whole, but lacks one or more moieties found in the whole. In some embodiments, a fragment consists of such a discrete portion. In some embodiments, a fragment consists of or comprises a characteristic structural element or moiety found in the whole.
  • a polymer fragment comprises or consists of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 or more monomeric units (e.g., residues) as found in the whole polymer.
  • monomeric units e.g., residues
  • a polymer fragment comprises or consists of at least about 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more of the monomeric units (e.g., residues) found in the whole polymer.
  • the whole material or entity may in some embodiments be referred to as the “parent” of the fragment.
  • homology refers to the overall relatedness between polymeric molecules, e.g., between polypeptide molecules.
  • polymeric molecules such as antibodies are considered to be “homologous” to one another if their sequences are at least 80%, 85%, 90%, 95%, or 99% identical.
  • polymeric molecules are considered to be “homologous” to one another if their sequences are at least 80%, 85%, 90%, 95%, or 99% similar.
  • a human is an embryo, a fetus, an infant, a child, a teenager, an adult, or a senior citizen.
  • Humanized as is known in the art, the term "humanized” is commonly used to refer to antibodies (or antibody components) whose amino acid sequence includes VH and VL region sequences from a reference antibody raised in a non-human species (e.g., a mouse), but also includes modifications in those sequences relative to the reference antibody intended to render them more "human-like” , i.e., more similar to human germline variable sequences.
  • a "humanized” antibody is one that immunospecifically binds to an antigen of interest and that has a framework (FR) region having substantially the amino acid sequence as that of a human antibody, and a complementary determining region (CDR) having substantially the amino acid sequence as that of a non-human antibody.
  • a humanized antibody comprises substantially all of at least one, and typically two, variable domains (Fab, Fab', F(ab')2, FabC, Fv) in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin (i.e., donor immunoglobulin) and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence.
  • a humanized antibody also comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin constant region.
  • a humanized antibody contains both the light chain as well as at least the variable domain of a heavy chain.
  • the antibody also may include a CHI , hinge, CH2, CH3, and, optionally, a CH4 region of a heavy chain constant region.
  • a humanized antibody only contains a humanized VL region.
  • a humanized antibody only contains a humanized VH region.
  • a humanized antibody contains humanized VH and VL regions.
  • Identity refers to the overall relatedness between polymeric molecules, e.g., between nucleic acid molecules e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules.
  • polymeric molecules are considered to be “substantially identical” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical.
  • Calculation of the percent identity of two nucleic acid or polypeptide sequences can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes).
  • the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or substantially 100% of the length of a reference sequence. The nucleotides at corresponding positions are then compared.
  • the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences.
  • the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller (CABIOS, 1989, 4: 11-17), which has been incorporated into the ALIGN program (version 2.0).
  • nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
  • the percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix.
  • an appropriate reference measurement may be or comprise a measurement in a particular system (e.g., in a single individual) under otherwise comparable conditions absent presence of (e.g., prior to and/or after) a particular agent or treatment, or in presence of an appropriate comparable reference agent.
  • an appropriate reference measurement may be or comprise a measurement in comparable system known or expected to respond in a particular way, in presence of the relevant agent or treatment.
  • Peptide refers to a polypeptide that is typically relatively short, for example having a length of less than about 100 amino acids, less than about 50 amino acids, less than about 40 amino acids less than about 30 amino acids, less than about 25 amino acids, less than about 20 amino acids, less than about 15 amino acids, or less than 10 amino acids.
  • composition refers to a composition in which an active agent is formulated together with one or more pharmaceutically acceptable carriers.
  • the active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population.
  • a pharmaceutical composition may be specially formulated for administration in a particular form (e.g., in a solid form or a liquid form), and/or may be specifically adapted for, for example: oral administration (for example, as a drenche [aqueous or non-aqueous solutions or suspensions], tablet, capsule, bolus, powder, granule, paste, etc, which may be formulated specifically for example for buccal, sublingual, or systemic absorption); parenteral administration (for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation, etc); topical application (for example, as a cream, ointment, patch or spray applied for example to skin, lungs, or oral cavity); intravaginal or intrarectal administration (for example, as a pessary, suppository, cream, or foam); ocular administration; nasal or pulmonary administration, etc.
  • oral administration for example, as a drenche [a
  • Polypeptide As used herein refers to a polymeric chain of amino acids.
  • a polypeptide has an amino acid sequence that occurs in nature.
  • a polypeptide has an amino acid sequence that does not occur in nature.
  • a polypeptide has an amino acid sequence that is engineered in that it is designed and/or produced through action of the hand of man.
  • a polypeptide may comprise or consist of natural amino acids, non-natural amino acids, or both.
  • a polypeptide may comprise or consist of only natural amino acids or only nonnatural amino acids.
  • a polypeptide may comprise D-amino acids, L- amino acids, or both.
  • a polypeptide may comprise only D-amino acids. In some embodiments, a polypeptide may comprise only L-amino acids. In some embodiments, a polypeptide may include one or more pendant groups or other modifications, e.g., modifying or attached to one or more amino acid side chains, at the polypeptide’s N-terminus, at the polypeptide’s C-terminus, or any combination thereof. In some embodiments, such pendant groups or modifications may be selected from the group consisting of acetylation, amidation, lipidation, methylation, pegylation, etc., including combinations thereof. In some embodiments, a polypeptide may be cyclic, and/or may comprise a cyclic portion.
  • a polypeptide is not cyclic and/or does not comprise any cyclic portion.
  • a polypeptide is linear.
  • a polypeptide may be or comprise a stapled polypeptide.
  • the term “polypeptide” may be appended to a name of a reference polypeptide, activity, or structure; in such instances it is used herein to refer to polypeptides that share the relevant activity or structure and thus can be considered to be members of the same class or family of polypeptides.
  • exemplary polypeptides within the class whose amino acid sequences and/or functions are known; in some embodiments, such exemplary polypeptides are reference polypeptides for the polypeptide class or family.
  • a member of a polypeptide class or family shows significant sequence homology or identity with, shares a common sequence motif (e.g., a characteristic sequence element) with, and/or shares a common activity (in some embodiments at a comparable level or within a designated range) with a reference polypeptide of the class; in some embodiments with all polypeptides within the class).
  • a member polypeptide shows an overall degree of sequence homology or identity with a reference polypeptide that is at least about 30-40%, and is often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more and/or includes at least one region (e.g., a conserved region that may in some embodiments be or comprise a characteristic sequence element) that shows very high sequence identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99%.
  • a conserved region that may in some embodiments be or comprise a characteristic sequence element
  • Such a conserved region usually encompasses at least 3-4 and often up to 20 or more amino acids; in some embodiments, a conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more contiguous amino acids.
  • a relevant polypeptide may comprise or consist of a fragment of a parent polypeptide.
  • a useful polypeptide as may comprise or consist of a plurality of fragments, each of which is found in the same parent polypeptide in a different spatial arrangement relative to one another than is found in the polypeptide of interest (e.g., fragments that are directly linked in the parent may be spatially separated in the polypeptide of interest or vice versa, and/or fragments may be present in a different order in the polypeptide of interest than in the parent), so that the polypeptide of interest is a derivative of its parent polypeptide.
  • Reference As used herein describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and/or determined substantially simultaneously with the testing or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and/or comparison to a particular possible reference or control.
  • Specific binding refers to an ability to discriminate between possible binding partners in the environment in which binding is to occur.
  • a binding agent that interacts with one particular target when other potential targets are present is said to "bind specifically" to the target with which it interacts.
  • specific binding is assessed by detecting or determining degree of association between the binding agent and its partner; in some embodiments, specific binding is assessed by detecting or determining degree of dissociation of a binding agent-partner complex; in some embodiments, specific binding is assessed by detecting or determining ability of the binding agent to compete an alternative interaction between its partner and another entity. In some embodiments, specific binding is assessed by performing such detections or determinations across a range of concentrations.
  • an agent when used herein with reference to an agent having an activity, is understood by those skilled in the art to mean that the agent discriminates between potential target entities or states. For example, an in some embodiments, an agent is said to bind “specifically” to its target if it binds preferentially with that target in the presence of one or more competing alternative targets. In many embodiments, specific interaction is dependent upon the presence of a particular structural feature of the target entity (e.g., an epitope, a cleft, a binding site). It is to be understood that specificity need not be absolute. In some embodiments, specificity may be evaluated relative to that of the binding agent for one or more other potential target entities (e.g., competitors).
  • specificity is evaluated relative to that of a reference specific binding agent. In some embodiments specificity is evaluated relative to that of a reference non-specific binding agent. In some embodiments, the agent or entity does not detectably bind to the competing alternative target under conditions of binding to its target entity. In some embodiments, binding agent binds with higher on-rate, lower off-rate, increased affinity, decreased dissociation, and/or increased stability to its target entity as compared with the competing alternative target(s).
  • Specificity is a measure of the ability of a particular ligand to distinguish its binding partner from other potential binding partners.
  • the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest.
  • One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result.
  • the term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.
  • Substantial identity refers to a comparison between amino acid or nucleic acid sequences. As will be appreciated by those of ordinary skill in the art, two sequences are generally considered to be “substantially identical” if they contain identical residues in corresponding positions. As is well known in this art, amino acid or nucleic acid sequences may be compared using any of a variety of algorithms, including those available in commercial computer programs such as BLAS TN for nucleotide sequences and BLASTP, gapped BLAST, and PSLBLAST for amino acid sequences. Exemplary such programs are described in Altschul et al., Basic local alignment search tool, J. Mol.
  • two sequences are considered to be substantially identical if at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more of their corresponding residues are identical over a relevant stretch of residues.
  • the relevant stretch is a complete sequence.
  • the relevant stretch is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 or more residues.
  • CDR In the context of a CDR, reference to "substantial identity” typically refers to a CDR having not more than a small number (e.g., 3, 2, or 1) an amino acid sequence changes relative to that of a reference CDR. In some embodiments, a CDR that is substantially identical to a reference CDR differs from that reference CDR by one or more amino acid changes at the end of the reference CDR; in some such embodiments, the relevant CDR is identical to the reference CDR other than at one or both ends. As is known in the art, CDR elements typically have a length within a range of a few amino acids (e.g., 3, 4, 5, 6, or 7) to about 20 or 30 amino acids (see, for example, Collis et al. J. Mol. Biol.
  • a CDR may be considered to be substantially identical to a reference CDR when it shares at least about 80% (or less for a shorter CDR), at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or at least about 100% identity with the reference CDR.
  • Substantial sequence homology is used herein to refer to a comparison between amino acid or nucleic acid sequences. As will be appreciated by those of ordinary skill in the art, two sequences are generally considered to be “substantially homologous” if they contain homologous residues in corresponding positions. Homologous residues may be identical residues. Alternatively, homologous residues may be non-identical residues will appropriately similar structural and/or functional characteristics. For example, as is well known by those of ordinary skill in the art, certain amino acids are typically classified as “hydrophobic” or “hydrophilic” amino acids., and/or as having “polar” or “non-polar” side chains. Substitution of one amino acid for another of the same type may often be considered a “homologous” substitution. Typical amino acid categorizations are summarized below:
  • amino acid or nucleic acid sequences may be compared using any of a variety of algorithms, including those available in commercial computer programs such as BLAS TN for nucleotide sequences and BLASTP, gapped BLAST, and PSL BLAST for amino acid sequences.
  • Exemplary such programs are described in Altschul, et al., Basic local alignment search tool, J. Mol. Biol., 215(3): 403-410, 1990; Altschul, et al., Methods in Enzymology; Altschul, et al., "Gapped BLAST and PSLBLAST: a new generation of protein database search programs", Nucleic Acids Res.
  • two sequences are considered to be substantially homologous if at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more of their corresponding residues are homologous over a relevant stretch of residues.
  • the relevant stretch is a complete sequence.
  • the relevant stretch is at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, at least 90, at least 95, at least 100, at least 125, at least 150, at least 175, at least 200, at least 225, at least 250, at least 275, at least 300, at least 325, at least 350, at least 375, at least 400, at least 425, at least 450, at least 475, at least 500 or more residues.
  • Treat As used herein, the term “treat,” “treatment,” or “treating” is used to refer to one or more of partial or complete alleviation, amelioration, relief, inhibition, prevention, delay of onset of, reduction in severity of and/or reduction in frequency (e.g., incidence) of one or more symptoms or features of a disease, disorder, and/or condition.
  • treatment may be prophylactic; for example may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition.
  • treatment may be administered to a subject who exhibits early signs of the disease, disorder, and/or condition, and may, for example, decrease risk of developing pathology associated with the disease, disorder, and/or condition and/or delay onset and/or decrease rate of development or worsening of one or more features of a disease, disorder and/or condition.
  • treatment refers to administration of a therapy that partially or completely alleviates, ameliorates, relieves, inhibits, delays onset of, reduces severity of, and/or reduces incidence of one or more symptoms, features, and/or causes of a particular disease, disorder, and/or condition.
  • such treatment may be of a subject who does not exhibit signs of the relevant disease, disorder and/or condition and/or of a subject who exhibits only early signs of the disease, disorder, and/or condition.
  • such treatment may be of a subject who exhibits one or more signs of the relevant disease, disorder and/or condition.
  • treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, and/or condition.
  • treatment may be of a subject known to have one or more susceptibility factors, e.g., that are statistically correlated with increased risk of development of the relevant disease, disorder, and/or condition.
  • treatment may be prophylactic; in some embodiments, treatment may be therapeutic.
  • variant refers to a molecule or entity (e.g., that are or comprise a nucleic acid, protein, or small molecule) that shows significant structural identity with a reference molecule or entity but differs structurally from the reference molecule or entity, e.g., in the presence or absence or in the level of one or more chemical moieties as compared to the reference molecule or entity. In some embodiments, a variant also differs functionally from its reference molecule or entity. In many embodiments, whether a particular molecule or entity is properly considered to be a “variant” of a reference is based on its degree of structural identity with the reference molecule.
  • a biological or chemical reference molecule in typically characterized by certain characteristic structural elements.
  • a variant, by definition, is a distinct molecule or entity that shares one or more such characteristic structural elements but differs in at least one aspect from the reference molecule or entity.
  • a polypeptide may have a characteristic sequence element comprised of a plurality of amino acids having designated positions relative to one another in linear or three-dimensional space and/or contributing to a particular structural motif and/or biological function;
  • a nucleic acid may have a characteristic sequence element comprised of a plurality of nucleotide residues having designated positions relative to one another in linear or three-dimensional space.
  • a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and/or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nucleic acid (e.g., that are attached to the polypeptide or nucleic acid backbone).
  • moieties e.g., carbohydrates, lipids, phosphate groups
  • a variant polypeptide or nucleic acid shows an overall sequence identity with a reference polypeptide or nucleic acid that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 99%.
  • a variant polypeptide or nucleic acid does not share at least one characteristic sequence element with a reference polypeptide or nucleic acid.
  • a reference polypeptide or nucleic acid has one or more biological activities.
  • a variant polypeptide or nucleic acid shares one or more of the biological activities of the reference polypeptide or nucleic acid.
  • a variant polypeptide or nucleic acid lacks one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid shows a reduced level of one or more biological activities as compared to the reference polypeptide or nucleic acid. In some embodiments, a polypeptide or nucleic acid of interest is considered to be a “variant” of a reference polypeptide or nucleic acid if it has an amino acid or nucleotide sequence that is identical to that of the reference but for a small number of sequence alterations at particular positions.
  • a variant polypeptide or nucleic acid comprises about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1 substituted residues as compared to a reference.
  • a variant polypeptide or nucleic acid comprises a very small number (e.g., fewer than about 5, about 4, about 3, about 2, or about 1) number of substituted, inserted, or deleted, functional residues (e.g., residues that participate in a particular biological activity) relative to the reference.
  • a variant polypeptide or nucleic acid comprises not more than about 5, about 4, about 3, about 2, or about 1 addition or deletion, and, in some embodiments, comprises no additions or deletions, as compared to the reference.
  • a variant polypeptide or nucleic acid comprises fewer than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and commonly fewer than about 5, about 4, about 3, or about 2 additions or deletions as compared to the reference.
  • a reference polypeptide or nucleic acid is one found in nature.
  • a reference polypeptide or nucleic acid is a human polypeptide or nucleic acid.
  • glycoengineered polypeptides and compositions comprising the same having the ability to degrade one or more IgG4 antibodies (e.g., IgG4 autoantibodies) by binding to IgG4 antibodies with a first moiety (e.g., an anti-IgG4 antibody) and binding to an endocytic receptor with a second moiety comprising one or more glycans, thus targeting the IgG4 antibody (or a complex thereof) for degradation.
  • IgG4 antibodies e.g., IgG4 autoantibodies
  • a first moiety e.g., an anti-IgG4 antibody
  • an endocytic receptor e.g., an anti-IgG4 antibody
  • a glycoengineered polypeptide is engineered by introduction of one or more glycosylation sites on a glycoengineered polypeptide, resulting in an engineered glycosylation profile that mediates endocytic receptor degradation of the glycoengineered polypeptides and the target to which it binds.
  • a glycoengineered polypeptide described herein 1) has homogeneous glycosylation; 2) can degrade large targets such as immune complexes; 3) has a defined ligand-to-antibody ratio; 4) has defined glycosylation sites; 5) can activate more diverse and powerful degradation receptors; and/or 6) can engage in protein degradation in a highly optimized manner.
  • a glycoengineered polypeptide may be employed as a novel therapeutic to treat autoimmune diseases, e.g., a disease associated with IgG4 autoantibodies such as Pemphigus Vulgaris/Folaceus; Muscle-specific kinase (MuSK) myasthenia gravis (MG); Idiopathic Thrombotic Thrombocytopenia Purpura (iTTP); idiopathic membranous nephropathy (iMN); Chronic Inflammatory Demyelinating Polyneuropathy (CIDP); or Immunoglobulin G4-Related Disease (IgG4-RD).
  • IgG4 autoantibodies such as Pemphigus Vulgaris/Folaceus; Muscle-specific kinase (MuSK) myasthenia gravis (MG); Idiopathic Thrombotic Thrombocytopenia Purpura (iTTP); idiopathic membranous nephropathy (iMN
  • IgG4 antibodies e.g., IgG4 autoantibodies
  • Immunoglobulin 4 is one of four human IgG subclasses and has several unique functional characteristics. IgG4 is also the least common of the 4 subclasses of IgG. It exhibits low affinity for complement and for most Fc receptors, see Huijbers MG et al., (2016) Annals of the NY Academy of Sciences, pp. 92-103, the entire contents of which are hereby incorporated by reference. IgG4 generally has a high affinity for its antigen, with binding occurring in a monovalent fashion, as IgG4 can exchange Fab-arms with other IgG4 molecules. Because of these characteristics, IgG4 is believed to block its targets and prevent inflammation, which, depending on the setting, can have a protective or pathogenic effect (Huijbers 2018).
  • IgG4-mediated autoimmune diseases include but are not limited to, thrombotic thrombocytopenic purpura, chronic inflammatory polyneuropathy, limbic encephalitis, neuromotonia, Morvan syndrome, pemphigus follaceus, pemphigus vulgaris, parasomnia, myasthenia gravis, and membranous nephropathy.
  • glycoengineered polypeptides comprising a first moiety that comprises an antibody agent that binds to an IgG4 antibody (e.g., an IgG4 autoantibody).
  • an IgG4 antibody e.g., an IgG4 autoantibody.
  • Exemplary anti-IgG4 antibodies that can be used in a first moiety of a glycoengineered polypeptide described herein are provided in Tables 1 and 2.
  • exemplary anti-IgG4 antibodies provided herein can be useful in treating and/or preventing diseases associated IgG4 autoantibodies, e.g., as disclosed herein.
  • exemplary anti-IgG4 antibodies disclosed herein preferentially bind to IgG4 with minimal or substantially no binding (e.g., no detectable binding) to non-IgG4 antibodies (e.g., antibodies of a different IgG subclass or antibodies of a different isotype).
  • an anti-IgG4 antibody disclosed herein binds to an IgG4 antibody (e.g., an IgG4 autoantibody) at one or more of: a CHI domain, a hinge domain, a CH2 domain or a CH3 domain.
  • an IgG4 antibody e.g., an IgG4 autoantibody
  • IgG4 polypeptide sequence is provided as SEQ ID NO: 201, corresponding to UniProt Accession Number P01861 : ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSV FLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNST YRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEM TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRW QEGNVFSCSVMHEALHNHYTQKSLSLSLELQLEESCAEAQDGELDGLWTTITIFITLFLLS VCYSATVTFF
  • An IgG4 polypeptide comprises the following domains: a CHI domain, and Ig-like 1 domain, a hinge domain, a CH2 domain, an Ig-like 2 domain, a CH3 domain and an Ig-like 3 domain.
  • a CHI domain comprises the sequence of
  • an Ig-like 1 domain comprises the sequence of
  • a CHI domain comprises the sequence of
  • a CHI domain comprises the sequence of
  • a CHI domain comprises the sequence of
  • a CHI domain comprises the sequence of
  • a hinge domain comprises the sequence of ESKYGPPCPSCP (SEQ ID NO: 204).
  • a CH2 domain comprises the sequence of
  • an Ig-like 2 domain comprises the sequence of
  • a CH3 domain comprises the sequence of GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLEL (SEQ ID NO: 207).
  • an Ig-like 3 domain comprises the sequence of PQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF FLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLS (SEQ ID NO: 208).
  • a first moiety of a glycoengineered polypeptide is or comprises an anti-IgG4 antibody agent. In some embodiments, a first moiety of a glycoengineered polypeptide is or comprises an antigen binding fragment of an anti-IgG4 antibody agent. In some embodiments, a first moiety of a glycoengineered polypeptide is or comprises a variable light (VL) polypeptide. In some embodiments, a first moiety of a glycoengineered polypeptide is or comprises a variable heavy (VH) polypeptide. In some embodiments, a first moiety of a glycoengineered polypeptide is or comprises a VL polypeptide and a VH polypeptide.
  • a first moiety of a glycoengineered polypeptide is or comprises a Fab fragment, a Fab’ fragment, a F(ab’)2 fragment, a Fd’ fragment, or a Fd fragment.
  • a first moiety of a glycoengineered polypeptide is or comprises a Fab fragment.
  • a first moiety of a glycoengineered polypeptide is or comprises an scFv.
  • a first moiety of a glycoengineered polypeptide is or comprises a single domain antibody (e.g., a VHH or a VNAR) or a fragment thereof.
  • a first moiety of a glycoengineered polypeptide is or comprises a diabody. In some embodiments, a first moiety of a glycoengineered polypeptide is or comprises a nanobody. In some embodiments, a first moiety of a glycoengineered polypeptide is or comprises an Fc fusion.
  • an antibody agent is a monoclonal or polyclonal antibody. In some embodiments, an antibody agent is a recombinant antibody. In some embodiments, an antibody agent is humanized, chimeric or fully human.
  • an anti-IgG4 antibody agent further comprises one or more additional antigen binding domains.
  • the one or more additional antigen binding domains bind to an antigen other than an Ig4 antibody (e.g., other than a CHI, hinge, CH2 and/or CH3 domain of an IgG4 antibody).
  • an anti-IgG4 antibody agent is a multi-specific antibody agent, e.g., a bispecific or a trispecific antibody agent).
  • an anti-IgG4 antibody agent further comprises one or more domains such as an extracellular domain, a hinge domain, a transmembrane domain and/or an intracellular domain.
  • an anti-IgG4 antibody agent is a chimeric receptor which binds to an IgG4 antibody (e.g., an IgG4 autoantibody).
  • glycoengineered polypeptides comprising: (a) a first moiety comprising one or more peptides that specifically binds to an IgG4 antibody (e.g., IgG4 autoantibody) or a fragment or a complex thereof; and (b) a second moiety comprising one or more glycans conjugated to the first moiety at one or more glycosylation sites.
  • IgG4 antibody e.g., IgG4 autoantibody
  • a second moiety comprising one or more glycans conjugated to the first moiety at one or more glycosylation sites.
  • a glycoengineered polypeptide disclosed herein comprises a first moiety, a second moiety and one or more additional elements.
  • a glycoengineered polypeptide comprises: an N-glycosylation site, a linker, a spacer, a signal peptide, a tag, a half-life extender or a combination thereof.
  • a glycoengineered polypeptide comprises one or more N- glycosylation sites in a first moiety.
  • a first moiety comprises one or more N-glycosylation sites that are naturally occurring and/or one or more N-glycosylation sites that are engineered into a first moiety.
  • an engineered N-glycosylation site (also referred to herein as a glycosite or a glycotag) is or comprises the sequence of GGGGANSTAPAPAPAHHHHHHHHHH (SEQ ID NO: 161).
  • an engineered N-glycosylation site is or comprises the sequence of GGGGANSTAPAPAPA (SEQ ID NO: 163).
  • an engineered N-glycosylation site is or comprises the sequence of GGGGANSTAPAPAPAPAPAPAGGGGANSTAPAPAPA (SEQ ID NO: 168).
  • an engineered N-glycosylation site is or comprises the sequence of GGGGANSTAPAPAPAPAPAPAPAPAANSTAPAPAPAHHHHAPAPAPA (SEQ ID NO: 169).
  • a glycoengineered polypeptide comprises a linker.
  • a linker comprises a Gly-Ser linker, or an EAAAK linker.
  • a linker comprises a (Gly-Gly-Gly-Gly-Ser)n linker, wherein n is an integer between 0 to 20.
  • a glycoengineered polypeptide comprises a spacer.
  • a spacer comprises one or more nucleotides which separates a first nucleic acid sequence from a subsequent nucleic acid sequence.
  • a spacer comprises a nucleic acid sequence which encodes one or more peptides that separates a first encoded polypeptide sequence from a subsequent encoded polypeptide sequence.
  • a glycoengineered polypeptide comprises a signal peptide, e.g., as disclosed herein.
  • a signal peptide is a native signal peptide.
  • a signal peptide is not a native signal peptide.
  • a signal peptide is derived from a Leishmania species. In certain embodiments, a signal peptide is derived from Leishmania tarentolae. In certain embodiments, a signal peptide is derived from Leishmania major.
  • the signal peptide is an invertase signal peptide derived from Leishmania tarentolae.
  • the signal peptide is an alkaline phosphatase signal peptide derived from Leishmania major.
  • a signal peptide comprises an amino acid sequence of SEQ ID NO: 164, or a portion thereof. In certain embodiments, a signal peptide comprises an amino acid sequence of SEQ ID NO: 165, or a portion thereof. In certain embodiments, a signal peptide comprises an amino acid sequence of SEQ ID NO: 166, or a portion thereof. In certain embodiments, a signal peptide comprises an amino acid sequence of SEQ ID NO: 162, or a portion thereof. In certain embodiments, a signal peptide is processed and removed from a glycoengineered polypeptide.
  • Exemplary signal peptide SPinv, a modified signal peptide from Leishmania tarentolae invertase, SEQ ID NO: 164: MIASSVRHAVILLLVAVAMMAAVIA.
  • Exemplary signal peptide SPinv, the native signal peptide from Leishmania tarentolae invertase, SEQ ID NO: 165: MIASSVRHAVILLLVAVAMMAAAVIA.
  • Exemplary signal peptide native signal peptide from Leishmania major alkaline phosphatase, SEQ ID NO: 166: MASRLVRVLAAAMLVAAAVS.
  • Exemplary signal peptide native signal peptide derived from Leishmania tarentolae invertase Spinv4, SEQ ID NO: 162: MIASSVRHAVILLLVAVAMMGGVIA.
  • a glycoengineered polypeptide comprises a tag.
  • a tag is a moiety that can be used for purifying and/or identifying a glycoengineered polypeptide disclosed herein.
  • a tag is not a glycotag.
  • a tag comprises a His tag, a Myc tag, or a GST tag.
  • a tag comprises a cleavable tag.
  • a tag is a His tag (HHHHHHHHHH; SEQ ID NO: 167).
  • a half-life extender comprises albumin or a fragment or a variant thereof.
  • a half-life extender comprises a Fc domain, e.g., with or without mutations in an Fc domain.
  • a glycoengineered polypeptide comprises a first moiety comprising one or more peptides that specifically binds to an IgG4 antibody (e.g., an IgG4 autoantibody) or a fragment thereof.
  • a glycoengineered polypeptide comprises a first moiety comprising one or more antibody agents that specifically binds to an IgG4 antibody (e.g., an IgG4 autoantibody) or a fragment thereof.
  • one or more antibody agents comprise an antigen binding fragment.
  • an antibody agent comprises a full antibody, a Fab fragment, an scFv, a nanobody, a duobody, or a single domain antibody (e.g., a VHH).
  • an antibody agent is or comprises a Fab fragment.
  • an antibody agent comprises a light chain polypeptide (e.g., a light chain variable region (VL)) and/or a heavy chain polypeptide (e.g.., a heavy chain variable region (VH)).
  • VL light chain variable region
  • VH heavy chain variable region
  • Light chain e.g., light chain variable region(VL) polypeptides
  • polypeptides comprising light chain (LC) sequences (e.g., light chain variable region sequence(s)) that, for example, may be useful as a first moiety in glycoengineered polypeptides described herein targeting hIgG4 antibodies (e.g., hIgG4 autoantibodies); in some such embodiments, such provided polypeptides are useful and/or included in such glycoengineered polypeptides as described herein.
  • LC light chain
  • a glycoengineered polypeptide comprises a first moiety comprising a LC polypeptide comprising one or more CDRs and/or one or more framework regions.
  • a LC polypeptide comprises a light chain variable region (VL) polypeptide.
  • a glycoengineered polypeptide comprises a first moiety comprising a LC polypeptide comprising at least one LC CDR provided in Table 1 or a sequence with at least 85% identity thereto.
  • a LC polypeptide comprises one, two or three LC CDRs (e.g., a LC CDR1, a LC CDR2 and/or a LC CDR3).
  • a LC polypeptide comprises a LC CDR1.
  • a LC polypeptide comprises a LC CDR2.
  • a LC polypeptide comprises a LC CDR3.
  • a LC polypeptide comprises a LC CDR1, a LC CDR2 and a LC CDR3.
  • a LC polypeptide having a LC CDR1, a LC CDR2 and a LC CDR3, e.g., in a first moiety of a glycoengineered polypeptide is capable of binding (e.g., specifically binding) to a hIgG4 antibody, e.g., a hIgG4 autoantibody.
  • a glycoengineered polypeptide comprises a first moiety comprising a LC polypeptide further comprises one or more framework regions, and/or a constant region.
  • a glycoengineered polypeptide comprises a first moiety comprising a LC polypeptide comprising a light chain constant region and/or a heavy chain constant region.
  • a LC polypeptide comprises a light chain constant region or a portion thereof, (e.g., a lambda light chain constant region or a variant or portion thereof; or a kappa light chain constant region or a variant or a portion thereof).
  • a LC polypeptide comprises a light chain comprising the sequence RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 215).
  • a LC polypeptide comprises a light chain comprising the sequence GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPS KQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:
  • a LC polypeptide comprises a light chain comprising the sequence GQPKAAPS VTLFPPS SEELQ ANKATLVCLISDF YPGAVTVAWKAD S SPVKAGVETTTPS KQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:
  • a LC polypeptide comprises a light chain comprising the sequence GQPKAAPS VTLFPPS SEELQ ANKATLVCLISDF YPGAVTVAWKAD S SPVKAGVETTTPS KQSNNKYAASSYLSLTPEQWKSHKSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:
  • a LC polypeptide comprises (i) an LC CDR1 sequence provided in Table 1, (ii) a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%identity to an LC CDR1 sequence provided in Table 1; or (iii) a sequence having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to an LC CDR1 sequence provided in Table 1
  • a LC polypeptide comprises (i) an LC CDR2 sequence provided in Table 1, (ii) a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity to an LC CDR2 sequence provided in Table 1; or (iii) a sequence having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to an LC CDR2 sequence provided in Table 1
  • a LC polypeptide comprises (i) an LC CDR3 sequence provided in Table 1, (ii) a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to an LC CDR3 sequence provided in Table 1; or (iii) a sequence having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to an LC CDR3 sequence provided in Table 1
  • a LC polypeptide comprises (i) an LC CDR1, LC CDR2, and LC CDR3 sequence provided in Table l;(ii) a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity to an LC CDR1, LC CDR2, and LC CDR3 sequence provided in Table 1; or (iii) a sequence having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to an LC CDR1, LC CDR2, and LC CDR3 sequence provided in Table 1.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 111, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 111 ; (ii) an LC CDR2 of SEQ ID NO: 125, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO:
  • an LC CDR2 of SEQ ID NO: 126 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a sequence having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 126; and/or (iii) an LC CDR3 of SEQ ID NO: 137, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO:
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 114, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 114; (ii) an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 112; (ii) an LC CDR2 of SEQ ID NO: 128, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 112; (ii) an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 112; (ii) an LC CDR2 of SEQ ID NO: 129, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 115, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 115; (ii) an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 112; (ii) an LC CDR2 of SEQ ID NO: 130, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 116; (ii) an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 116; (ii) an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 116; (ii) an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 116; (ii) an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 117, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 117; (ii) an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 117, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 117; (ii) an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 118, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 118; (ii) an LC CDR2 of SEQ ID NO: 132, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 118, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 118; (ii) an LC CDR2 of SEQ ID NO: 133, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 119, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 119; (ii) an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 119, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 119; (ii) an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 120, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 120; (ii) an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 119, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 119; (ii) an LC CDR2 of SEQ ID NO: 135, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO:
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO:
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO:
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO:
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 152, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 152; (ii) an LC CDR2 of SEQ ID NO: 153, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 199, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 199; (ii) an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 119, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 119; (ii) an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a LC polypeptide comprises the sequence of SEQ ID NO: 85, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 85.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 86, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 86.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 87, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 87.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 88, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 88.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 89, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 89.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 90, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 90.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 91, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 91.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 92, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 92.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 93, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 93.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 94, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 94.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 95, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 95.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 96, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 96.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 97, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 97.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 98, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 98.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 99, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 99.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 100, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 100.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 101, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 101.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 102, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 102.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 103, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 103.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 104, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 104.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 105, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 105.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 106, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 106.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 107, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 107.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 108, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 108.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 109, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 109.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 110, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 110.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 195, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 195.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 191, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 191.
  • a LC polypeptide comprises the sequence of SEQ ID NO: 155, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 155.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 157, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 157.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 158, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 158.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a LC polypeptide comprises the sequence of SEQ ID NO: 160, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 160.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 85 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 85 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 94 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 94 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 87 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 87 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 88 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 88 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 89 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 89 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 86 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a LC polypeptide comprising a VL sequence of SEQ ID NO: 86 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises a LC polypeptide comprising a CDR1 having the consensus sequence RASQX1IX2X3X4LX5 (SEQ ID NO: 170), wherein Xi is chosen from T, S, or G; X2 is chosen from S or R; X3 is chosen from S or K; X4 is chosen from Y or F, and X5 is chosen from N, S, or A.
  • a first moiety of a glycoengineered polypeptide comprises a LC polypeptide comprising a CDR2 having the consensus sequence X1X2SX3LX4X5, (SEQ ID NO: 171), wherein Xi is chosen from A, D, or G; X2 is chosen from A, T, or G; X3 is chosen from S, T, or M; X4 is chosen from L or Q; and X5 is chosen from S or A.
  • a first moiety of a glycoengineered polypeptide comprises a LC polypeptide comprising a CDR3 having the consensus sequence QQX1X2X3X4PX5T (SEQ ID NO: 209), wherein Xi is chosen from S, T, or L; X2 is chosen from Y or N; X3 is chosen from S, or N; and X4 is chosen from S, T, A, P or Y; and X5 is chosen from L or F.
  • a first moiety of a glycoengineered polypeptide comprises a LC polypeptide comprising a CDR1 having the consensus sequence SGSSSNIGHNYVXi (SEQ ID NO: 172), wherein Xi is chosen from N or A.
  • a first moiety of a glycoengineered polypeptide comprises a LC polypeptide comprising a CDR3 having the consensus sequence GSYDXiSGSTRV (SEQ ID NO: 173), wherein Xi is chosen from L, I, N, T or S.
  • a first moiety of a glycoengineered polypeptide comprises a LC polypeptide comprising a CDR1 having the consensus sequence RSSX1SLX2X3SDGX4TYLX5 (SEQ ID NO: 174), wherein Xi is chosen from Q, L, or H; X2 is chosen from V or L; X3 is chosen from Y or F; X4 is chosen from N or H; and X5 is chosen from N, H, or I.
  • a first moiety of a glycoengineered polypeptide comprises a LC polypeptide comprising a CDR2 having the consensus sequence XiVSNRDS (SEQ ID NO: 175), wherein Xi is chosen from K or E.
  • a first moiety of a glycoengineered polypeptide comprises a LC polypeptide comprising a CDR3 having the consensus sequence X1QX2X3X4X5PX6T (SEQ ID NO: 176), wherein Xi is chosen from M or Q; X2 is chosen from G, S, T, or L; X3 is chosen from S, T, Y, or N; X4 is chosen from H, S, or N; X5 is chosen from W, S, T, A, Y, or P; and Xe is chosen from P, I, L, or F.
  • Heavy chain (e.g., heavy chain variable region(VH)) polypeptides e.g., heavy chain variable region(VH) polypeptides
  • HC heavy chain
  • VH heavy chain variable region
  • a glycoengineered polypeptide comprises a first moiety comprising a HC polypeptide comprising at least one HC CDR of an anti-IgG4 antibody agent as provided in Table 2 or a sequence with at least 85% identity thereto.
  • an HC polypeptide comprises one, two or three HC CDRs (e.g., an HC CDR1, an HC CDR2 and/or an HC CDR3).
  • an HC polypeptide comprises an HC CDR1.
  • an HC polypeptide comprises an HC CDR2.
  • an HC polypeptide comprises an HC CDR3.
  • an HC polypeptide comprises an HC CDR1, an HC CDR2 and an HC CDR3.
  • a glycoengineered polypeptide comprises a first moiety comprising a HC polypeptide comprising an HC CDR1, an HC CDR2 and an HC CDR3 is capable of binding (e.g., specifically binding) to hIgG4 antibody, e.g., a hIgG4 autoantibody.
  • a glycoengineered polypeptide comprises a first moiety comprising a HC polypeptide further comprises one or more framework regions, and/or a heavy chain constant region, or a portion or a variant thereof (e.g., a CHI, CH2 and/or CH3 region).
  • an HC polypeptide comprises a CHI, a CH2 or a CH3 or a combination thereof.
  • an HC polypeptide comprises a CH2 and CH3, e.g., an Fc domain.
  • a Fc domain comprises a mammalian Fc domain.
  • a Fc domain comprises a dog, a cat, a mouse, a rat, a rabbit, a primate or a human Fc domain.
  • a Fc domain comprises a human Fc domain.
  • a Fc domain comprises a dog Fc domain.
  • a Fc domain comprises a cat Fc domain.
  • an Fc domain is chosen from an Fc domain of an immunoglobulin isotype.
  • an immunoglobulin isotype comprises IgA, IgD, IgG, IgM, or IgE.
  • an Fc domain comprises an Fc domain of an IgG, e.g., a human IgG.
  • an IgG is or comprises IgGl, lgG2, lgG3, or lgG4.
  • an Fc region is a wildtype Fc region, e.g., a wildtype human Fc region.
  • an Fc region comprises a variant, e.g., an Fc region comprising an addition, substitution, or deletion of at least one amino acid residue in an Fc region which results in, e.g., reduced or ablated affinity for at least one Fc receptor.
  • the Fc region of an antibody interacts with a number of receptors or ligands including Fc Receptors (e.g., FcyRI, FcyRIIA, FcyRIIIA), the complement protein Clq, and other molecules such as proteins A and G.
  • Fc Receptors e.g., FcyRI, FcyRIIA, FcyRIIIA
  • the complement protein Clq e.g., FcyRI, FcyRIIA, FcyRIIIA
  • ADCC antibody dependent cell-mediated cytotoxicity
  • ADCP Antibody-dependent cellular phagocytosis
  • CDC complement dependent cytotoxicity
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 33; (ii) an HC CDR2 of SEQ ID NO: 45, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 33; (ii) an HC CDR2 of SEQ ID NO: 45, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 33; (ii) an HC CDR2 of SEQ ID NO: 46, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 33; (ii) an HC CDR2 of SEQ ID NO: 47, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 33; (ii) an HC CDR2 of SEQ ID NO: 45, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 33; (ii) an HC CDR2 of SEQ ID NO: 45, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 33; (ii) an HC CDR2 of SEQ ID NO: 45, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 34, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 34; (ii) an HC CDR2 of SEQ ID NO: 48, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 34, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 34; (ii) an HC CDR2 of SEQ ID NO: 48, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 34, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 34; (ii) an HC CDR2 of SEQ ID NO: 48, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 34, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 34; (ii) an HC CDR2 of SEQ ID NO: 48, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 34, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 34; (ii) an HC CDR2 of SEQ ID NO: 48, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 192, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 192; (ii) an HC CDR2 of SEQ ID NO: 193, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 35, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 35; (ii) an HC CDR2 of SEQ ID NO: 49, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 35, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 35; (ii) an HC CDR2 of SEQ ID NO: 49, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 36, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 36; (ii) an HC CDR2 of SEQ ID NO: 50, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 36, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 37; (ii) an HC CDR2 of SEQ ID NO: 50, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 37, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 36; (ii) an HC CDR2 of SEQ ID NO: 50, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 36, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 36; (ii) an HC CDR2 of SEQ ID NO: 50, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 38, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 38; (ii) an HC CDR2 of SEQ ID NO: 51, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 38, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 38; (ii) an HC CDR2 of SEQ ID NO: 52, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 39, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 39; (ii) an HC CDR2 of SEQ ID NO: 51, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 38, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 38; (ii) an HC CDR2 of SEQ ID NO: 52, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 40, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 40; (ii) an HC CDR2 of SEQ ID NO: 53, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 41, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 41; (ii) an HC CDR2 of SEQ ID NO: 54, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 42, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 42; (ii) an HC CDR2 of SEQ ID NO: 55, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 43, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 43; (ii) an HC CDR2 of SEQ ID NO: 56, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 42, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 42; (ii) an HC CDR2 of SEQ ID NO: 57, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 42, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 42; (ii) an HC CDR2 of SEQ ID NO: 58, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 42, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 42; (ii) an HC CDR2 of SEQ ID NO: 59, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 44, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 44; (ii) an HC CDR2 of SEQ ID NO: 60, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 196, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 196; (ii) an HC CDR2 of SEQ ID NO: 197, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 42, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 42; (ii) an HC CDR2 of SEQ ID NO: 188, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least
  • a HC polypeptide comprises: (i) an HC CDR1 of SEQ ID NO: 149, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto or a having at least 5 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 149; (ii) an HC CDR2 of SEQ ID NO: 150, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at
  • a HC polypeptide comprises the sequence of SEQ ID NO: 1, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 1.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 2, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 2.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 3, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 3.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a HC polypeptide comprises the sequence of SEQ ID NO: 4, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 4.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a HC polypeptide comprises the sequence of SEQ ID NO: 5, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 5.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a HC polypeptide comprises the sequence of SEQ ID NO: 6, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 6.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 7, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 7.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 8, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 8.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 9, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 9.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 10, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 10.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a HC polypeptide comprises the sequence of SEQ ID NO: 11, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 11.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 12, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 12.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 13, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 13.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 14.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 15, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 15.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 16, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 16.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 17, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 17.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 18, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 18.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 19, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 19.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 20, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 20.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 21, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 21.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 22, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 22.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 23, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 23.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 24, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 24.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 25, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 25.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 26, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 26.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 27, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 27.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 28, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 28.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 29, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 29.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 30, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 30.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 31, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 31.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 32, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 32.
  • a HC polypeptide comprises the sequence of SEQ ID NO: 194, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 194.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a HC polypeptide comprises the sequence of SEQ ID NO: 190, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 190.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a HC polypeptide comprises the sequence of SEQ ID NO: 154, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 154.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a HC polypeptide comprises the sequence of SEQ ID NO: 156, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 156.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a HC polypeptide comprises the sequence of SEQ ID NO: 159, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity thereto, or a sequence having at least 5, 10, or 20 alterations (e.g., substitutions, deletions or insertions (e.g., conservative substitutions)) relative to SEQ ID NO: 159.
  • substitutions, deletions or insertions e.g., conservative substitutions
  • a first moiety of a glycoengineered polypeptide comprises: a HC polypeptide comprising a VH sequence of SEQ ID NO: 1 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a HC polypeptide comprising a VH sequence of SEQ ID NO: 2 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a HC polypeptide comprising a VH sequence of SEQ ID NO: 3 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a HC polypeptide comprising a VH sequence of SEQ ID NO: 17 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a HC polypeptide comprising a VH sequence of SEQ ID NO: 16 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a HC polypeptide comprising a VH sequence of SEQ ID NO: 13 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR1 having the consensus sequence X1YX2MX3 (SEQ ID NO: 177), wherein Xi is chosen from T or D; X2 is chosen from A, S, or T; and X3 is chosen from S or N.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR2 having the consensus sequence
  • XIX2X3X 4 X5X6X 7 X 8 X9XIOYXI 1X12X13 X14KX15 (SEQ ID NO: 178), wherein Xi is chosen from A, Y, D, or S; X2 is chosen from I or V; X3 is chosen from S or Y; X4 is chosen from G, S, Y, or H; X5 is chosen from S or R; Xe is chosen from G or S; X7 is chosen from D, R, T, or S; X 8 is chosen from N, T, or Y; X9 is chosen from T, I, H, or Y; X10 is chosen from Y, F, or is absent; Xu is A or N; X12 is D, P, A, or S; X13 is A or S; X14 is V or L; and X15 is G or S.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR3 having the consensus sequence X1DWNFX2DX3 (SEQ ID NO:
  • Xi is chosen from M or L; X2 is chosen from F or H; and X3 is chosen from Y, F, and M.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR1 having the consensus sequence X1X2TFSX3X4 (SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR3 having the consensus sequence MTX1WTLDX2 (SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR1 having the consensus sequence X1YX2MN (SEQ ID NO: 182), wherein Xi is chosen from D or T; and X2 is chosen from S or T.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR3 having the consensus sequence SLGX1PHDX2 (SEQ ID NO: 183), wherein Xi is chosen from Y or F; and X2 is chosen from Y or G.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR1 having the consensus sequence SXiAIS (SEQ ID NO: 184), wherein Xi is chosen from A or Y.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR2 having the consensus sequence GIIPXiFGEAAYAQKFQG (SEQ ID NO: 185), wherein Xi is chosen from E or V.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR1 having the consensus sequence SSRYYWXi (SEQ ID NO: 186), wherein Xi is chosen from G, D, or A.
  • a first moiety of a glycoengineered polypeptide comprises a HC polypeptide comprising a CDR3 having the consensus sequence X1GAX2X3EGDY (SEQ ID NO: 187), wherein Xi is chosen from E or D; X2 is chosen from S or E; and X3 is chosen from P or V.
  • a glycoengineered polypeptide comprises a first moiety comprising a light chain comprising a variable region comprising one, two or three LC CDRs and a heavy chain comprising a variable region comprising one, two or three HC CDRs.
  • a glycoengineered polypeptide comprises a first moiety comprising a light chain comprising a LC CDR1, a LC CDR2 and a LC CDR3, and a heavy chain comprising an HC CDR1, an HC CDR2 and HC CDR3.
  • the first moiety of a glycoengineered polypeptide comprising a light chain comprising an LC CDR1, a LC CDR2 and a LC CDR3, and a heavy chain comprising an HC CDR1, an HC CDR2 and HC CDR3 is able to specifically bind to a IgG4 antibody (e.g., an IgG4 autoantibody) or a fragment thereof.
  • a IgG4 antibody e.g., an IgG4 autoantibody
  • a first moiety of a glycoengineered polypeptide comprises a VL sequence provided in Table 1 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5, 10, or 20 substitutions relative to a VL sequence provided in Table 1; and a VH sequence provided in Table 2 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to a VH sequence provided in Table 2.
  • a first moiety of a glycoengineered polypeptide comprises one, two, or three LC CDRs provided in Table 1 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto; and one, two, or three HC CDRs provided in Table 2, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto.
  • a first moiety of a glycoengineered polypeptide comprises: (a) a light chain comprising: (i) an LC CDR1 provided in Table 1 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to an LC CDR1 provided in Table 1; (ii) an LC CDR2 provided in Table 1 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 111, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC CDR2 of SEQ ID NO: 125, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 111, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC CDR2 of SEQ ID NO: 125, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 111, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC CDR2 of SEQ ID NO: 125, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC
  • an LC CDR3 of SEQ ID NO: 136 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 136; and (ii) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 33; an HC CDR2 of SEQ ID NO: 46, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 111, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC CDR2 of SEQ ID NO: 125, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC
  • an LC CDR3 of SEQ ID NO: 136 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 136; and (ii) an HC CDR1 of SEQ ID NO: 33, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 33; an HC CDR2 of SEQ ID NO: 47, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 111, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC CDR2 of SEQ ID NO: 125, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 111, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC CDR2 of SEQ ID NO: 125, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 111, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC CDR2 of SEQ ID NO: 125, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 111 ; an LC
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 113, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 113; an LC CDR2 of SEQ ID NO: 127, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 113, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 113; an LC CDR2 of SEQ ID NO: 127, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 114, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 114; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 128, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of
  • an LC CDR3 of SEQ ID NO: 137 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 137; and (ii) an HC CDR1 of SEQ ID NO: 35, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 35; an HC CDR2 of SEQ ID NO: 49, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 129, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 129, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • an LC CDR3 of SEQ ID NO: 139 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 139; and (ii) an HC CDR1 of SEQ ID NO: 35, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 35; an HC CDR2 of SEQ ID NO: 49, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 115, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 115; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 115; an LC CDR
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 130, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 130;
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 112, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2 of SEQ ID NO: 126, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 112; an LC CDR2
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR
  • an LC CDR3 of SEQ ID NO: 140 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 140; and (ii) an HC CDR1 of SEQ ID NO: 36, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 36; an HC CDR2 of SEQ ID NO: 50, or a sequence with at least 85%,
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR
  • an LC CDR3 of SEQ ID NO: 140 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 140; and (ii) an HC CDR1 of SEQ ID NO: 36, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 36; an HC CDR2 of SEQ ID NO: 50, or a sequence with at least 85%,
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR
  • an LC CDR3 of SEQ ID NO: 142 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 142; and (ii) an HC CDR1 of SEQ ID NO: 36, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 36; an HC CDR2 of SEQ ID NO: 50, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 116, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 116; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 117, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 117; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 117, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 117; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 117; an LC CDR
  • an LC CDR3 of SEQ ID NO: 144 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 144; and (ii) an HC CDR1 of SEQ ID NO: 38, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 38; an HC CDR2 of SEQ ID NO: 52, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 117, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 117; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 117; an LC CDR
  • an LC CDR3 of SEQ ID NO: 143 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 143; and (ii) an HC CDR1 of SEQ ID NO: 38, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 38; an HC CDR2 of SEQ ID NO: 51, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 117, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 117; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 117, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 117; an LC CDR2 of SEQ ID NO: 131, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 118, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 118; an LC CDR2 of SEQ ID NO: 132, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 118; an LC CDR
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 118, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 118; an LC CDR2 of SEQ ID NO: 133, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 118; an LC CDR
  • an LC CDR3 of SEQ ID NO: 145 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 145; and (ii) an HC CDR1 of SEQ ID NO: 40, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 40; an HC CDR2 of SEQ ID NO: 53, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 119, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 119; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 119, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 119; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 120, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 120; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 120; an LC CDR2 of S
  • an LC CDR3 of SEQ ID NO: 148 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 148; and (ii) an HC CDR1 of SEQ ID NO: 43, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 43; an HC CDR2 of SEQ ID NO: 56, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 119, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 119; an LC CDR2 of SEQ ID NO: 135, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 119; an LC CDR2
  • an LC CDR3 of SEQ ID NO: 148 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 148; and (ii) an HC CDR1 of SEQ ID NO: 42, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 42; an HC CDR2 of SEQ ID NO: 57, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 121, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 121; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 121; an LC CDR
  • an LC CDR3 of SEQ ID NO: 148 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 148; and (ii) an HC CDR1 of SEQ ID NO: 42, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 42; an HC CDR2 of SEQ ID NO: 57, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 122, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 122; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 123, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 123; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 123; an LC CDR
  • an LC CDR3 of SEQ ID NO: 147 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 147; and (ii) an HC CDR1 of SEQ ID NO: 42, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 42; an HC CDR2 of SEQ ID NO: 59, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 124, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 124; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 124; an LC CDR
  • an LC CDR3 of SEQ ID NO: 147 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 147; and (ii) an HC CDR1 of SEQ ID NO: 44, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 44; an HC CDR2 of SEQ ID NO: 60, or
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 199, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 199; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 199; an LC CDR
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 119, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 119; an LC CDR2 of SEQ ID NO: 134, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) an LC CDR1 of SEQ ID NO: 152, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 152; an LC CDR2 of SEQ ID NO: 153, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5 substitutions relative to SEQ ID NO: 152; an LC CDR
  • a first moiety of a glycoengineered polypeptide comprises a light chain polypeptide (LC polypeptide) as described herein.
  • a LC polypeptide comprises a light chain comprising the sequence RTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 215).
  • a LC polypeptide comprises a light chain comprising the sequence GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPS KQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:
  • a LC polypeptide comprises a light chain comprising the sequence
  • GQPKAAPS VTLFPPS SEELQ ANKATLVCLISDF YPGAVTVAWKAD S SPVKAGVETTTPS KQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:
  • a LC polypeptide comprises a light chain comprising the sequence GQPKAAPS VTLFPPS SEELQ ANKATLVCLISDF YPGAVTVAWKAD S SPVKAGVETTTPS KQSNNKYAASSYLSLTPEQWKSHKSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises a heavy chain polypeptide (HC polypeptide) as described herein.
  • a HC polypeptide comprises a CHI domain as described herein.
  • a CHI domain comprises the sequence of ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC (SEQ ID NO: 214).
  • a CHI domain comprises the sequence of ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSNFGTQTYTCNVDHKPSNTKVDKTVE (SEQ ID NO: 216).
  • a CHI domain comprises the sequence of ASTKGPSVFPLAPCSRSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYTCNVNHKPSNTKVDKRVE (SEQ ID NO: 217).
  • a CHI domain comprises the sequence of ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTKTYTCNVDHI ⁇ PSNTI ⁇ VDI ⁇ RVE (SEQ ID NO: 218).
  • a first moiety of a glycoengineered polypeptide comprises a light chain polypeptide (LC polypeptide) as described herein and a heavy chain polypeptide (HC polypeptide) as described herein.
  • LC polypeptide light chain polypeptide
  • HC polypeptide heavy chain polypeptide
  • a first moiety of a glycoengineered polypeptide comprises a VL sequence provided in Table 1 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto or a sequence having at least 5, 10, or 20 substitutions relative to a VL sequence provided in Table 1; and a VH sequence provided in Table 2 or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to a VH sequence provided in Table 2.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 85, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 85; and the sequence of SEQ ID NO: 1, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 1.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 85, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 85; and the sequence of SEQ ID NO: 2, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 2.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 85, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 85; and the sequence of SEQ ID NO: 3, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 3.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 85, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 85; and the sequence of SEQ ID NO: 4, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 4.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 85, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 85; and the sequence of SEQ ID NO: 5, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 5.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 85, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 85; and the sequence of SEQ ID NO: 6, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 6.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 85, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 85; and the sequence of SEQ ID NO: 7, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 7.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 86, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 86; and the sequence of SEQ ID NO: 8, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 8.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 86, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 86; and the sequence of SEQ ID NO: 9, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 9.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 86, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 86; and the sequence of SEQ ID NO: 10, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 10.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 86, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 86; and the sequence of SEQ ID NO: 11, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 11.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 86, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 86; and the sequence of SEQ ID NO: 12, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 12.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 87, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 87; and the sequence of SEQ ID NO: 13, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 13.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 87, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 87; and the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 14.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 88, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 88; and the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 14.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 89, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 89; and the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 14.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 90, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 90; and the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 14.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 91, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 91; and the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 14.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 91, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 91; and the sequence of SEQ ID NO: 15, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 15.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 90, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 90; and the sequence of SEQ ID NO: 15, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 15.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 92, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 92; and the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 14.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 93, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 93; and the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 14.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 86, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 86; and the sequence of SEQ ID NO: 14, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 14.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 94, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 94; and the sequence of SEQ ID NO: 16, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 16.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 95, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 95; and the sequence of SEQ ID NO: 17, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 17.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 94, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 94; and the sequence of SEQ ID NO: 18, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 18.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 94, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 94; and the sequence of SEQ ID NO: 19, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 19.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 96, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 96; and the sequence of SEQ ID NO: 17, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 17.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 97, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 97; and the sequence of SEQ ID NO: 17, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 17.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 98, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 98; and the sequence of SEQ ID NO: 20, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 20
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 98, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 98; and the sequence of SEQ ID NO: 21, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 21
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 99, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 99; and the sequence of SEQ ID NO: 20, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 20.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 99, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 99; and the sequence of SEQ ID NO: 22, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 22.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 98, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 98; and the sequence of SEQ ID NO: 23, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 23
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 100, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 100; and the sequence of SEQ ID NO: 24, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 24.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 101, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 101; and the sequence of SEQ ID NO: 24, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 24.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 102, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 102; and the sequence of SEQ ID NO: 24, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 24
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 103, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 103; and the sequence of SEQ ID NO: 25, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 25
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 104, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 104; and the sequence of SEQ ID NO: 26, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 26
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 105, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 105; and the sequence of SEQ ID NO: 27, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 27
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 106, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 106; and the sequence of SEQ ID NO: 28, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 28
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 107, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 107; and the sequence of SEQ ID NO: 29, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 29
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 108, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 108; and the sequence of SEQ ID NO: 30, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 30
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 109, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 109; and the sequence of SEQ ID NO: 31, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 31
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 110, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 110; and the sequence of SEQ ID NO: 32, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 32.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 195, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 195; and the sequence of SEQ ID NO: 194, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 191, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 191; and the sequence of SEQ ID NO: 190, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO:
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 94, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 94; and the sequence of SEQ ID NO: 17, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 17.
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 155, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 155; and the sequence of SEQ ID NO: 154, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 155; and the sequence of
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 157, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 157; and the sequence of SEQ ID NO: 156, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 157; and the sequence of
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 158, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 158; and the sequence of SEQ ID NO: 156, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 158; and the sequence of
  • a first moiety of a glycoengineered polypeptide comprises: (i) the sequence of SEQ ID NO: 160, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 160; and the sequence of SEQ ID NO: 159, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% thereto, or a sequence having at least 5, 10, or 20 substitutions relative to SEQ ID NO: 160; and the sequence of SEQ ID
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 1 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 85 or a sequence with a t least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 1 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 85 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 2 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 85 or a sequence with a t least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 2 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 85 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 3 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 85 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 3 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 85 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 17 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 94 or a sequence with a t least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 17 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 94 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 16 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 94 or a sequence with a t least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 16 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 94 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 13 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 87 or a sequence with a t least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 13 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 87 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 87 or a sequence with a t least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 87 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 88 or a sequence with a t least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 88 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 89 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 89 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 86 or a sequence with at least 85% identity thereto and a constant region of SEQ ID NO: 215 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a first moiety of a glycoengineered polypeptide comprises: (i) a HC polypeptide comprising a VH sequence of SEQ ID NO: 14 or a sequence with at least 85% thereto and a constant region of SEQ ID NO: 214 or a sequence with at least 85% thereto; and (ii) a LC polypeptide comprising a VL sequence of SEQ ID NO: 86 or a sequence with a t least 85% identity thereto and a constant region of SEQ ID NO: 220 or a sequence with at least 85% thereto.
  • a first moiety (e.g., a HC polypeptide and/or a LC polypeptide) further comprises the sequence of any one of SEQ ID NOs: 161, 163, 168 or 169. In some embodiments, a first moiety further comprises a sequence of any one of SEQ ID NOs: 165, 164, 166 or 162.
  • a glycoengineered polypeptide disclosed herein comprises a first moiety that specifically binds to an IgG4 antibody or a fragment or a complex thereof; and a second moiety comprising one or more glycans conjugated to the first moiety at one or more glycosylation sites.
  • glycan engagement with endocytic carbohydrate binding proteins and receptors enables different biological pathways. These essential biological pathways are involved in modulating immune responses, mediating protein clearance, protein turnover, and controlling trafficking of soluble glycoproteins, glycolipids and any natural molecule containing a glycan moiety.
  • the glycan-receptor interaction is determined by the glycan structure.
  • Glycan binding receptors are highly diverse and can be exploited by glycoengineering to develop novel therapeutics based on the concept of gly can-mediated protein degradation to treat different diseases, which include but are not limited to autoimmune disorders as disclosed herein.
  • a glycoengineered polypeptide comprising a second moiety having one or more glycans, as described herein is expected to activate natural degradation pathways.
  • a second moiety of a glycoengineered polypeptide disclosed herein comprises one or more glycans and specifically binds to one or more endocytic receptors.
  • Endocytic receptors as described herein capture glycoproteins via specific glycan structures to mediate degradation, e.g., lysosomal degradation.
  • Endocytic receptors are ubiquitous in human and can be found on different cells.
  • an endocytic receptor is or comprises an endocytic lectin.
  • the endocytic receptor is chosen from: an asialoglycoprotein receptor (ASGPR); a mannose binding receptor, a Cluster of Differentiation 206 (CD206) receptor; a DC-SIGN (Cluster of Differentiation 209 or CD209) receptor; a C-Type Lectin Domain Family 4 Member G (LSECTin) receptor; a macrophage inducible Ca2+-dependent lectin receptor (Mincle); a L- SIGN CD209L receptor; dectin- 1; dectin -2, langerin, macrophage mannose 2 receptor, BDCA- 2, DCIR, MBL, MDL, MICE, CLEC2, DNGR1, CLEC12B, DEC-205, CLEC10, and mannose 6 phosphate receptor (M6PR), or a combination thereof.
  • ASGPR asialoglycoprotein receptor
  • CD206 Cluster of Differentiation 206
  • a glycoengineered polypeptide comprising a first moiety that specifically binds to a target protein (e.g., an IgG4 autoantibody) and a second moiety comprising a glycan comprising terminal GlcNAc.
  • a target protein e.g., an IgG4 autoantibody
  • a second moiety comprising a glycan comprising terminal GlcNAc.
  • a glycoengineered polypeptide comprising a first moiety that specifically binds to a target protein (e.g., an IgG4 autoantibody) and a second moiety comprising a glycan comprising terminal GalNAc.
  • a target protein e.g., an IgG4 autoantibody
  • a second moiety comprising a glycan comprising terminal GalNAc.
  • a glycoengineered polypeptide comprising a first moiety that specifically binds to a target protein (e.g., an IgG41 autoantibody) and a second moiety comprising a glycan comprising terminal Gal.
  • a target protein e.g., an IgG41 autoantibody
  • a second moiety comprising a glycan comprising terminal Gal.
  • a glycoengineered polypeptide provided herein can comprise (i) a binding specificity to one or more target protein(s) (e.g., one or more IgG4 autoantibodies) and (ii) one or more N-glycan(s) with binding specificities to one or more endocytic receptor(s).
  • a glycoengineered polypeptide comprises one type of N-glycan with binding specificity to one type of endocytic receptor.
  • a glycoengineered polypeptide comprises one or more N- glycosylation sites in a first moiety. In some embodiments, one or more N-glycosylation sites in a first moiety are native N-glycosylation sites. In some embodiments, one or more N- glycosylation sites in a first moiety are engineered N-glycosylation sites. In some embodiments, a glycoengineered polypeptide comprises one or more native N-glycosylation sites and one or more engineered N-glycosylation sites.
  • a second moiety comprising one or more glycans is conjugated, e.g., linked, to a first moiety at one or more N-glycosylation sites.
  • a glycoengineered polypeptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more N-glycosylation sites (or glycosites; such as an N-glycosylation consensus sequence). These N-glycosylation sites can be glycosylated by an N-glycan such that the resulting glycoengineered bifunctional binding protein can engage with or bind to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more endocytic receptor molecules.
  • a glycoengineered polypeptide comprises two types of N-glycans with binding specificities to two different endocytic receptors.
  • a glycoengineered polypeptide provided herein can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more polypeptide chains. Each chain can be produced in a different cell line.
  • the glycoengineered polypeptide can be an antibody and one type of N-glycan is on the Fc domain and another type of N-glycan is on the Fab domain (e.g., the variable regions) of the antibody.
  • a glycoengineered polypeptide comprises: (i) a first type of N- glycan with binding specificity to a first endocytic receptor wherein the first type of N-glycan is present at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more glycosites thus engaging with or binding to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more molecular of the first endocytic carbohydrate-binding protein or receptor ; and (ii) a second type of N-glycan with binding specificity to a second endocytic receptor wherein the second N-glycan is present at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more glycosites so that a single bifunctional binding protein can engage with or bind to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more molecules of the second endocytic receptor(s).
  • a glycoengineered polypeptide comprises: (i) a first type of N- glycan with binding specificity to a first endocytic receptor wherein the first type of N-glycan is present at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more glycosites thus engaging with or binding to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more molecular of the first endocytic receptor ; (ii) a second type of N- glycan with binding specificity to a second endocytic receptor wherein the second N-glycan is present at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more glycosites so that a single bifunctional binding protein can engage with or bind to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more molecules of the second endocytic receptor(s); and (iii) a third type of N-glycan with binding specificity to a third endocytic receptor wherein the third N-glycan is present at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10
  • a glycoengineered polypeptide provided herein has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more glycosites.
  • at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% of the glycosites in the population at one specific position are glycosylated.
  • At least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% of the glycosites in the population are glycosylated.
  • N-glycans that can be present at the glycosites of the glycoengineered polypeptide provided herein are described herein.
  • a glycosite is an N-glycosylation consensus sequence.
  • the consensus sequence can be N-X-S/T, or N-X-C, wherein X is any amino acid except proline.
  • a glycosite is or comprises the sequence of GGGGANSTAPAPAPA (SEQ ID NO: 163).
  • a glycosite is or comprises the sequence of GGGGANSTAPAPAPAHHHHHHHHHH (SEQ ID NO: 161).
  • a glycosite is or comprises the sequence of GGGGANSTAPAPAPAPAPAPAGGGGANSTAPAPAPA (SEQ ID NO: 168). [0501] In some embodiments, a glycosite is or comprises the sequence of GGGGANSTAPAPAPAPAPAPAPAANSTAPAPAPAHHHHHHAPAPA (SEQ ID NO: 169).
  • an N-glycan is conjugated to the glycoengineered polypeptide at at least one, two, three, or four N-glycosylation sites.
  • an N-glycan is conjugated to the glycoengineered polypeptide at one, two, three, or four N-glycosylation sites.
  • an N-glycosylation site is naturally occurring.
  • an N-glycosylation site is engineered into the amino acid sequence of the first moiety.
  • one or more of the N-glycosylation sites are engineered into the amino acid sequence of the first moiety of the glycoengineered polypeptide (e.g., one or more of the N-glycosylation sites are not present in a wild-type, or naturally occurring form of the first moiety). In certain embodiments, at least one of the N-glycosylation sites is engineered into the amino acid sequence of the first moiety of the glycoengineered polypeptide. In certain embodiments, at least two of the N-glycosylation sites are engineered into the amino acid sequence of the first moiety of the glycoengineered polypeptide.
  • At least three of the N-glycosylation sites are engineered into amino acid sequence of the first moiety of the glycoengineered polypeptide. In certain embodiments, at least four of the N-glycosylation sites are engineered into the amino acid sequence of the first moiety of the glycoengineered polypeptide. In certain embodiments, one or more of the engineered N-glycosylation sites are glycotags fused to the N- and/or C-terminus of the amino acid sequence of the first moiety of the glycoengineered polypeptide via a peptide linker. In certain embodiments, a glycotag is fused to the N-terminus of first moiety of the glycoengineered polypeptide.
  • a glycotag is fused to the C-terminus of first moiety of the glycoengineered polypeptide. In certain embodiments, a glycotag is fused to the N- and the C-terminus of first moiety of the glycoengineered polypeptide. In certain embodiments, one or more of the N-glycosylation sites are natural N-glycosylation sites (e.g., one or more of the N-glycosylation sites are present in a wild-type, or naturally occurring form of the first moiety). In certain embodiments, at least one of the N-glycosylation sites is a natural N-glycosylation site. In certain embodiments, at least two of the N-glycosylation sites are natural N-glycosylation sites.
  • a glycoengineered polypeptide that specifically binds to a target protein associated with a disease (e.g., an IgG4 autoantibody), comprising a first moiety and a second moiety.
  • a glycoengineered polypeptide comprising a first moiety that specifically binds to a target protein associated with a disease (e.g., an IgG4 autoantibody), and a second moiety that binds specifically to an endocytic receptor, wherein the second moiety comprises a glycan structure.
  • a glycoengineered polypeptide comprising a first moiety that specifically binds to a target protein and a second moiety comprising an N- glycan selected from the group consisting of GlcNAc2Man3GlcNAc2, GalNAc2GlcNAc2Man3 GlcNAc2, Gal2GlcNAc2Man3GlcNAc2, Man3 GlcNAc, GlcNAc lMan3 GlcNAc2, Gal2GlcNAc2Man3 GlcNAc2, Gal 1 GlcNAc2Man3 GlcNAc2, GalNAc 1 GlcNAc2Man3 GlcNAc2, GlcNAc3Man3 GlcNAc2, GlcNAc4Man3 GlcNAc2, Gal3GlcNAc3Man3 GlcNAc2, GalNAc3 GlcNAc3Man3 GlcNAc2, GalNAc4GlcNAc4Man3 GlcNAc2, Gal3GlcNA
  • increasing the number of glycan structures on a glycoengineered polypeptide increases the rate of lysosomal degradation as compared to an otherwise similar glycoengineered polypeptide with fewer glycan structures.
  • the number of glycan structures on a glycoengineered polypeptide disclosed herein is 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more or 10 or more glycan structures.
  • a glycoengineered polypeptide disclosed herein comprises a glycan structure having a monoantennary structure.
  • a glycoengineered polypeptide disclosed herein comprises a glycan structure having a biantennary structure.
  • a glycoengineered polypeptide disclosed herein comprises a glycan structure having a triantennary structure.
  • a glycoengineered polypeptide disclosed herein comprises a glycan structure having a tetraantennary structure.
  • the glycan structure comprises a biantennary structure.
  • the glycan structure comprises a biantennary GalNAc.
  • the biantennary GalNac binds to an asialoglycoprotein receptor (ASGPR) or a fragment or variant thereof, or a complex comprising ASGPR.
  • ASGPR asialoglycoprotein receptor
  • the N-glycan has a structure of: wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N-acetylglucosamine (GlcNAc) residue and the black circle represents a mannose (Man) residue, and wherein X represents an amino acid residue of the first moiety.
  • the N-glycan specifically binds to one or more endocytic receptors, e.g., that mediate lysosomal degradation. In some embodiments, the N-glycan specifically binds to ASGPR.
  • the endocytic receptor is or comprises ASGPR or a fragment or variant thereof, or a complex comprising ASGPR.
  • the glycan structure of the second moiety comprises a terminal GalNac.
  • ASGPR-mediated degradation in the hepatocyte has many applications.
  • ASGPR binding to the N-glycan structure disclosed herein can result in the selective degradation of one or more target proteins (e.g., an IgG4 autoantibody).
  • target proteins e.g., an IgG4 autoantibody.
  • ASGPR-mediated degradation can lead to removal of cytokines, chemokines and hormones.
  • ASGPR-mediated degradation can be used for the delivery of the target molecules to the hepatocyte endosome.
  • ASGPR-mediated degradation is applicable for various diseases, while limiting systemic toxicity.
  • the 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more N-glycosylation sites can be glycosylated by the N-glycan such that the resulting glycoengineered polypeptide can engage with or bind to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more endocytic receptor molecules.
  • the glycoengineered polypeptide is glycosylated by the N-glycan at at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 of the N- glycosylation sites.
  • the glycoengineered polypeptide is glycosylated by the N-glycan at at least 2 N-glycosylation sites.
  • the glycoengineered polypeptide is glycosylated by the N-glycan at at least 3 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at at least 4 N- glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at at least 5 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at at least 6 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at at least 7 N- glycosylation sites.
  • the glycoengineered polypeptide is glycosylated by the N-glycan at at least 8 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at at least 9 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at at least 10 N- glycosylation sites.
  • the glycoengineered polypeptide is glycosylated by the N-glycan at 2 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at 3 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at 4 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at 5 N- glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at 6 N-glycosylation sites.
  • the glycoengineered polypeptide is glycosylated by the N-glycan at 7 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at 8 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at 9 N- glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated by the N-glycan at 10 N-glycosylation sites. In certain embodiments, the glycoengineered polypeptide is glycosylated at an Asn amino acid residue of the glycoengineered polypeptide.
  • the N-glycosylation site is an N-glycosylation consensus sequence. In certain embodiments, the N-glycosylation site comprises a consensus sequence of N-X-S/T or N- X-C, wherein X is any amino acid except proline.
  • At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95, or at least 98% of the N- glycosylation sites are occupied by an N-glycan.
  • At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95, or at least 98% of the N-glycosylation sites are occupied by an N-glycan of the structure: linked to the glycoengineered polypeptide at one or more N-glycosylation sites, wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N- acetylglucosamine (GlcNAc) residue the black circle represents a mannose (Man) residue, and X represents an amino acid residue of the glycoengineered polypeptide .
  • GalNAc N-acetyl galactosamine
  • Man mannose
  • X represents an amino acid residue of the glycoengineered polypeptide .
  • At least 10% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 20% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 30% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 40% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 50% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 60% of the N-glycosylation sites are occupied by the N-glycan.
  • At least 70% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 80% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 90% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 95% of the N-glycosylation sites are occupied by the N-glycan. In certain embodiments, at least 98% of the N-glycosylation sites are occupied by the N-glycan.
  • the N-glycan is linked to the glycoengineered polypeptide at at least one N-glycosylation site. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at at least two N-glycosylation sites. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at one, two, three, or four N-glycosylation sites. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at one N-glycosylation site. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at two N-glycosylation sites.
  • the N-glycan is linked to the glycoengineered polypeptide at three N-glycosylation sites. In certain embodiments, the N- glycan is linked to the glycoengineered polypeptide at four N-glycosylation sites. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at an Asn amino acid residue of the glycoengineered polypeptide. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at an N-glycosylation consensus sequence. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at a consensus sequence of N-X-S/T or N-X-C, wherein X is any amino acid except proline.
  • the glycoengineered polypeptide comprises two different N- glycans (e.g., a first and a second N-glycan), wherein each N-glycan is independently linked to the glycoengineered polypeptide at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more N-glycosylation sites, and wherein one of the N-glycans (e.g., the first N-glycan) has the structure: wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N-acetylglucosamine (GlcNAc) residue the black circle represents a mannose (Man) residue, and X represents an amino acid residue of the glycoengineered polypeptide.
  • N-glycans e.g., a first and a second N-glycan
  • the different N-glycans specifically bind to different endocytic receptors.
  • the first N-glycan specifically binds to ASGPR.
  • the other N-glycan is an N-glycan described in PCT7EP2022/057556, which is incorporated herein by reference in its entirety.
  • the first N-glycan is larger than the second N-glycan.
  • the first N-glycan is smaller than the second N-glycan.
  • the N-glycosylation sites predominantly or exclusively occupied by the larger N-glycan are more sterically accessible than the N- glycosylation sites predominantly or exclusively occupied by the smaller N-glycan.
  • the other N-glycan is A2. In certain embodiments, the other N-glycan is AlGalNAcl or A2GalNAcl . In certain embodiments, the N-glycans are linked to the glycoengineered polypeptide at an Asn amino acid residue of the glycoengineered polypeptide. In certain embodiments, the N-glycans are linked to the glycoengineered polypeptide at an N- glycosylation consensus sequence. In certain embodiments, the N-glycans are linked to the glycoengineered polypeptide at a consensus sequence of N-X-S/T or N-X-C, wherein X is any amino acid except proline.
  • the first N-glycan is linked to the glycoengineered polypeptide at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more N-glycosylation sites
  • the second N-glycan is linked to the glycoengineered polypeptide at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more N-glycosylation sites.
  • the glycoengineered polypeptide further comprises a third N- glycan, wherein the third N-glycan is linked to the glycoengineered polypeptide at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more N-glycosylation sites.
  • the third N-glycan specifically binds to a different endocytic receptor than the first and/or second N-glycan.
  • the third N-glycan is an N-glycan described in PCI7EP2022/057556, which is incorporated herein by reference in its entirety.
  • the third N-glycan is A2.
  • the third N-glycan is Al GalNAcl or A2GalNAcl.
  • the third N-glycan is linked to the glycoengineered polypeptide at an Asn amino acid residue of the glycoengineered polypeptide. In certain embodiments, the third N-glycan is linked to the glycoengineered polypeptide at an N-glycosylation consensus sequence. In certain embodiments, the third N-glycan is linked to the glycoengineered polypeptide at a consensus sequence of N-X-S/T or N-X-C, wherein X is any amino acid except proline.
  • the second and/or third N-glycan specifically bind to an endocytic lectin.
  • the endocytic lectin is a mannose binding receptor.
  • the endocytic lectin is a Cluster of Differentiation 206 (CD206) receptor.
  • the endocytic lectin is a DC-SIGN (Cluster of Differentiation 209 or CD209) receptor.
  • the endocytic lectin is a C-Type Lectin Domain Family 4 Member G (LSECTin) receptor.
  • the endocytic lectin is a macrophage inducible Ca 2+ -dependent lectin receptor (Mincle).
  • the endocytic receptor is L-SIGN CD209L.
  • the endocytic receptor is asialoglycoprotein (ASGPR).
  • the endocytic receptor is dectin- 1.
  • the endocytic receptor is dectin-2.
  • the endocytic receptor is langerin.
  • the second and/or third N-glycan specifically bind to a receptor selected from the group consisting of macrophage mannose 2 receptor, BDCA-2, DCIR, MBL, MDL, MICE, CLEC2, CLEC10, DNGR1, CLEC12B, DEC-205, and mannose 6 phosphate receptor (M6PR).
  • a receptor selected from the group consisting of macrophage mannose 2 receptor, BDCA-2, DCIR, MBL, MDL, MICE, CLEC2, CLEC10, DNGR1, CLEC12B, DEC-205, and mannose 6 phosphate receptor (M6PR).
  • CD206 is a C-type lectin and phagocytic/endocytic recycling and signaling receptor. CD206 is expressed primarily by M2 anti-inflammatory macrophages, dendritic cells, and live sinusoidal endothelial cells.
  • DC-SIGN is a non-recycling, signaling receptor that targets both the ligand and receptor to the lysosome for degradation.
  • LSECTin is expressed on liver sinusoidal endothelial cells.
  • the glycoengineered polypeptide is glycosylated at two or more N-glycosylation sites by an N-glycan of the structure: wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N-acetylglucosamine (GlcNAc) residue the black circle represents a mannose (Man) residue, and X represents an amino acid residue of the glycoengineered polypeptide , and wherein two of the N-glycosylation sites are separated by at least 5 , at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 amino acids.
  • GalNAc N-acetyl galactosamine
  • Man mannose
  • X represents an amino acid residue of the glycoengineered polypeptide
  • the N-glycan is linked to the glycoengineered polypeptide at two N-glycosylation sites separated by a distance of about 5-10, about 10-20, about 20-30, about 30-40, about 40-50, about 50-60, about 60-70, about 70-80, about 80-90, about 90-100, about 100-150, about 150-200, or about 200-300 amino acids.
  • the amino acid separation between the N-glycosylation sites is the number of amino acids between the terminal amino acids of the N-glycosylation consensus sequence.
  • the glycoengineered polypeptide folds in space and, thus, has a three-dimensional geometry in addition to its primary amino acid structure.
  • this three-dimensional geometry, including the position of the N-glycan is not static but dynamic (see, for example, Re, S., et al Biophysical Reviews, 4, 179-187 (2012)).
  • the distance between N-glycosylation sites and/or N- glycans on a glycoengineered polypeptide may be from an equilibrium geometry of the glycoengineered polypeptide , as determined by any standard means known in the art, including for example computational modelling studies.
  • the N-glycan is linked to the glycoengineered polypeptide at two N-glycosylation sites separated by a distance of at least 1.0 nm.
  • the N-glycan is linked to the glycoengineered polypeptide at two N-glycosylation sites separated by a distance of about 1.0-5.0 nm. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at two N-glycosylation sites separated by a distance of about 1.5-3.0 nm. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at two N-glycosylation sites separated by a distance of about 1.5-
  • the N-glycan is linked to the glycoengineered polypeptide at three N-glycosylation sites each separated by a distance of about 1.0-5.0 nm. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at three N-glycosylation sites each separated by a distance of about 1.5-3.0 nm. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at three N-glycosylation sites each separated by a distance of about 1.5-2.5 nm. In certain embodiments, the N-glycans are separated by a distance of at least 1.0 nm. In certain embodiments, the N-glycans are separated by a distance of about 1.0 to about 5.0 nm. In certain embodiments, the N-glycans are separated by a distance of about
  • the distance between the N-glycosylation sites and/or N-glycans is chosen to minimize steric hindrance, for example between the glycoengineered polypeptide (s), the target protein(s), and/or the ASGPR receptor(s).
  • the distance between the N-glycosylation sites and/or N-glycans is chosen based on the separation of ASGPR receptors on a cell surface.
  • the distance between the N-glycosylation sites and/or N-glycans is chosen to be similar (e.g. no more than twice, or no less than half) to the separation of ASGPR receptors on a cell surface.
  • the N-glycan is linked to the glycoengineered polypeptide at an Asn amino acid residue of the glycoengineered polypeptide. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at an N-glycosylation consensus sequence. In certain embodiments, the N-glycan is linked to the glycoengineered polypeptide at a consensus sequence of N-X-S/T or N-X-C, wherein X is any amino acid except proline.
  • the present disclosure provides nucleic acid sequences encoding glycoengineered polypeptides as described herein.
  • a nucleic acid sequence is or comprises single stranded DNA (e.g., as in certain viral vectors).
  • a nucleic acid is or comprises double stranded DNA (e.g., as in certain viral vectors and/or certain plasmids).
  • a nucleic acid is or comprises RNA (e.g., as in certain viral vectors and/or as in mRNA therapeutics), etc.
  • Nucleic acids encoding glycoengineered polypeptides may be modified to include codons that are optimized for expression in a particular cell type (e.g., a Leishmania cell) or organism. Codon optimized sequences are synthetic sequences, and preferably encode an identical polypeptide (or biologically active fragment of a full length polypeptide which has substantially the same activity as the full length polypeptide) encoded by a non-codon optimized parent polynucleotide.
  • a coding region of a nucleic acids encoding glycoengineered polypeptides described herein, in whole or in part, may include an altered sequence to optimize codon usage for a particular cell type (e.g., a eukaryotic or prokaryotic cell).
  • a coding sequence for an antibody agent e.g., antigen binding fragment
  • an antibody agent e.g., antigen binding fragment
  • the coding sequence may be optimized for expression in a bacterial cells.
  • the coding sequence may be optimized for expression in a mammalian cell (e.g., a CHO cell).
  • a sequence may be described as a codon-optimized sequence.
  • Nucleic acid constructs of the present disclosure may be inserted into an expression vector or viral vector by methods known to the art, and nucleic acids may be operably linked to an expression control sequence.
  • a vector comprising any nucleic acids or fragments thereof described herein is further provided by the present disclosure. Any nucleic acids or fragments thereof described herein can be cloned into any suitable vector and can be used to transform or transfect any suitable host (e.g., Leishmania host cell). Selection of vectors and methods to construct them are commonly known to persons of ordinary skill in the art.
  • a vector may be or comprise a non-viral vector (e.g. a lipid nanoparticles or liposome).
  • nucleic acids and vectors of the present disclosure are isolated and/or purified.
  • the present disclosure also provides a composition comprising an isolated or purified nucleic acid, optionally in the form of a vector.
  • Isolated nucleic acids and vectors may be prepared using standard techniques known in the art including, for example, alkali/SDS treatment, CsCl binding, column chromatography, agarose gel electrophoresis, and/or other techniques well known in the art.
  • the composition can comprise other components as described further herein.
  • any method known to one skilled in the art for the insertion of nucleic acids into a vector may be used to construct expression vectors encoding a glycoengineered polypeptide described herein under control of transcriptional and/or translational control signals.
  • These methods may include in vitro recombinant DNA and synthetic techniques and in vivo recombination (see, e.g., Sambrook et al., Molecular Cloning, a Laboratory Manual, 2d edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989); and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and John Wiley & Sons, New York, N.Y. (1994), each of which is hereby incorporated by reference in its entirety).
  • Glycoengineered polypeptides disclosed herein can be useful for treating and/or preventing any disease or disorder in which a reduction in IgG4 antibody (e.g., IgG4 autoantibody) levels would be beneficial, e.g., to treat the disease, to prevent the disease, and/or to ameliorate one or more symptoms of the disease.
  • IgG4 antibody e.g., IgG4 autoantibody
  • PV Pemphigus vulgaris
  • PF pemphigus foliaceus
  • the pathophysiology of both PV and PF is associated with loss of function of desmosomes due to autoantibodies to desmoglein 1 (Dsgl) and desmoglein 3 (Dsg3). Because desmosomes bind keratinocytes together to form skin, disruption of desmosome function results in blistering, scaling, and erythematous patches.
  • Dsgl desmoglein 1
  • Dsg3 desmoglein 3
  • Desmosomes are cell-to-cell adhesion molecules which aid in epithelium formation by riveting keratinocytes together. Desmosomes are composed of networks of cadherin proteins, including desmoglein, a transmembrane glycoprotein which forms the extracellular core of the desmosome. Anti- desmoglein antibodies are thought to disturb the structural stability of the desmosome and thus the fidelity of the epithelium, leading to the skin and mucosal symptoms observed in PF and PV.
  • Dsgl autoantibodies typically dominate, whereas in PV, Dsg3 autoantibodies typically dominate.
  • PV patients that present with dermal but not oral symptoms may show a higher ratio of Dsgl to Dsg3 autoantibodies.
  • PV patients with severe symptom presentation may have both types of autoantibodies (Melchionda & Harman. Clin. Exp. Derm., 2019, 44(7): 740-746). In both cases, the disruption of desmosome function causes keratinocytes to split from one another, resulting in blister formation and lesions.
  • a glycoengineered polypeptide disclosed herein or a compositions comprising the same binds to one or more autoantibodies, e.g., one or more IgG4 autoantibodies, associated with pemphigus (e.g., PV or PF).
  • an autoantibody associated with pemphigus is an anti-Dsgl autoantibody (e.g., an IgG4 anti-Dsgl autoantibody).
  • an autoantibody associated with pemphigus is an anti-Dsg3 autoantibody (e.g., an IgG4 anti-Dsg3 autoantibody).
  • administration of a glycoengineered polypeptide disclosed herein or a compositions comprising the same to a subject having or identified with pemphigus results in a reduction in the level and/or activity of one or more autoantibodies (e.g., one or more IgG4 autoantibodies) in a subject.
  • a reduction in the level and/or activity of one or more autoantibodies is compared to (1) a healthy subject, or (2) the same subject prior to administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same.
  • a glycoengineered polypeptide disclosed herein comprises a first moiety that binds to an IgG4 antibody.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • an IgG4 antibody is an anti-desmoglein 1 (Dsgl) autoantibody or a fragment or a complex thereof.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • Dsg3 anti-desmoglein 3
  • Muscle-specific kinase Muscle-specific kinase (MuSK) myasthenia gravis (MG) [MuSK-MG]
  • MG Myasthenia gravis
  • AChR muscle acetylcholine receptor
  • MuSK is a transmembrane protein found in muscle which supports the contraction of muscles triggered by acetylcholine receptor activation. Activation of MuSK can lead to clustering of AChRs in the muscle via interactions with collagen Q and LRP4.
  • the extracellular domain of MuSK comprises three immunoglobulin-like regions to which anti-MuSK antibodies bind.
  • Anti-MuSK antibodies are typically of the IgG4 class. The binding of antibodies to the immunoglobulin-like regions of MuSK has an inhibitory effect on interactions between MuSK and collagen Q and LRP4, thereby disrupting AChR clustering and normal muscle function.
  • MG e.g., MuSK-MG
  • Available treatment options for patients with MG include administration of rituximab or the AChE inhibitor pyridostigmine, general immunosuppressant agents, intravenous immunoglobulin, plasmapheresis, thymectomy, or any combination thereof.
  • these treatments often confer little benefit in MuSK-MG, and none of the available treatment options can reduce the levels of IgG4 antibodies or immune complexes comprising the same.
  • a glycoengineered polypeptide disclosed herein or a composition comprising the same binds to one or more autoantibodies, e.g., one or more IgG4 autoantibodies, associated with MuSK-MG.
  • an autoantibody associated with MuSK-MG is anti-MuSK autoantibody (e.g., an IgG4 anti-MuSK autoantibody).
  • an autoantibody associated with MuSK-MG is an anti-nicotinic acetylcholine receptor (nAChR) IgG4 autoantibody (e.g., an IgG4 anti-nAChR autoantibody).
  • an autoantibody associated with MuSK-MG is an anti-low-density lipoprotein receptor-related protein 4 (LRP4) IgG4 autoantibody (e.g., an IgG4 anti-LRP4 autoantibody).
  • LRP4 low-density lipoprotein receptor-related protein 4
  • administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same to a subject having or identified with MuSK-MG results in a reduction in the level and/or activity of one or more autoantibodies (e.g., one or more IgG4 autoantibodies) in a subject.
  • a reduction in the level and/or activity of one or more autoantibodies is compared to (1) a healthy subject, or (2) the same subject prior to administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same.
  • a glycoengineered polypeptide disclosed herein comprises a first moiety that binds to an IgG4 antibody.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • nAchR non-nicotinic acetylcholine receptor
  • an IgG4 antibody e.g., IgG4 autoantibody
  • MoSK muscle-specific kinase
  • an IgG4 antibody (e.g., IgG4 autoantibody) is an an anti-low-density lipoprotein receptor-related protein 4 (LRP4) autoantibody or a fragment or a complex thereof.
  • an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-titin autoantibody or a fragment or a complex thereof.
  • an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-ryanodine receptor autoantibody or a fragment or a complex thereof.
  • an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-agrin autoantibody or a fragment or a complex thereof. In some embodiments, an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-collagen Q autoantibody or a fragment or a complex thereof. In some embodiments, an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-K v 1.4 potassium channel autoantibody or a fragment or a complex thereof. In some embodiments, an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-cortactin autoantibody or a fragment or a complex thereof.
  • TTP Thrombotic Thrombocytopenia Purpura
  • Thrombotic thrombocytopenic purpura is a life-threatening rare disorder characterized by a deficiency in the activity of ADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13) (Ercig B. et al. J. Biol. Chem., 2021).
  • ADAMTS13 is a mettaloprotease that cleaves the von Willebrand factor (VWF) thus preventing accumulation of ultralarge VWF (ULVWF) multimers.
  • iTTP autoimmune nature - immune TTP
  • autoantibodies targeting ADAMTS13 cause loss of ADAMTS13 activity and/or promote ADAMTS13 polypeptide clearance resulting in accumulation of highly pro- thrombotic ULVWF multimers (Ercig B et al 2021).
  • iTTP has an annual incidence rate of 1.5 to 6 cases per million adults per year.
  • the acute phase of iTTP often presents itself with purpura, fever, neurological manifestations, renal dysfunctions, hemolytic anemia with schistocytes, and thrombocytopenia.
  • VWF When enzymatically active, ADAMTS13 cleaves VWF or ULVWF (e.g., on the surface of endothelial cells).
  • VWF is produced by vascular endothelium and megakaryocytes. VWF circulates in a globular form; however, exposure to shear forces and binding to exposed collagen at the site of a damaged vessel promotes its unfolding. VWF recruits platelets to sites of vascular injury. The ability of VWF to bind circulating platelets is dependent on its multimeric size, with largest multimers being most potent in capturing platelets during primary hemostasis. Cleavage of VWF serves to reduce the size of VWF polymers in circulation.
  • VWF polymers Under normal physical conditions, the multimeric size of VWF is controlled by ADAMTS13. In the absence of a functional ADAMTS13 polypeptide, VWF polymers are not adequately processed, resulting in spontaneous adhesion of blood platelets, which presents as severe, life-threatening microvascular thrombosis (Ercig B. et al. J. Biol. Chem., 2021).
  • ADAMTS13 polypeptides circulate in a closed conformation. Upon binding to VWF, ADAMTS13 polypeptides adopt a short-lived transient open conformation that allows for interaction with VWF.
  • ADAMTS13 polypeptides may adopt an open conformation.
  • anti-ADAMST13 autoantibodies can induce a conformational change in ADAMTS13 resulting in exposure of one or more neoepitopes (Roose E et al., (2020) vol. 136, number 3). It has been reported that in some cases of iTTP, open conformation ADAMTS13 precedes a drop in ADAMTS13 activity thus suggesting that open conformation AD AMTS 13 may be useful as a biomarker for iTTP.
  • conformational changes of ADAMTS13 polypeptide may result in exposure of neoepitopes that may trigger the development of autoantibodies.
  • an open ADAMTS13 polypeptide conformation can be used as a biomarker for iTTP.
  • anti- D AMTS 13 autoantibodies can change the conformation of AD AMTS 13 from a closed conformation to an open conformation.
  • TTP TTP
  • iTTP TTP-specific triglyceride
  • iTTP TTP-specific triglyceride
  • iTTP plasma exchange and/or immunosuppressive and/or administration of a steroids, an anti-CD20 antibody, an anti- vWF antibody, or a combination thereof.
  • none of the available treatment options reduce and/or remove anti-ADAMTS13 autoantibodies or immune complexes comprising the same (Ercig B. et al. J. Biol. Chem., 2021).
  • Anti-ADAMTS13 autoantibodies Anti-ADAMTS13 autoantibodies
  • iTTP may be induced by autoimmunity to ADAMTS13 (e.g., by the development of anti- ADAMTS13 autoantibodies) (Roose 2020; Ercig 2021).
  • Anti-ADAMTS13 autoantibodies may inhibit ADAMTS13 activity and/or clear ADAMTS13 from circulation.
  • an anti- AD AMTS 13 autoantibody or a fragment or a complex thereof is characterized in that it binds to a ADAMTS13 polypeptide or a variant or a complex thereof.
  • anti- AD AMTS 13 autoantibodies or immune complexes comprising the same inhibit ADAMTS13 polypeptide activity.
  • binding of an anti-ADAMTS13 autoantibody to an ADAMTS13 polypeptide or a variant or fragment thereof induces a conformational change in an ADAMTS13 polypeptide or a variant or fragment thereof.
  • binding of an anti- AD AMTS 13 autoantibody to an AD AMTS 13 polypeptide or a variant or fragment thereof alters a level and/or activity of ADAMTS13. In some embodiments, binding of an anti- AD AMTS 13 autoantibody to an ADAMTS13 reduces the amount of ADAMTS13. In some embodiments, binding of an anti- AD AMTS 13 autoantibody to an AD AMTS 13 increase the amount of AD AMTS 13 in an open conformation. In some embodiments, binding of an anti- AD AMTS 13 autoantibody to an ADAMTS13 reduces the activity of ADAMTS13.
  • AD AMTS 13 Deficiency in AD AMTS 13 may result in accumulation of ULVWF multimers, which spontaneously bind to platelets and may lead to formation of microthrombi that obstruct the microvasculature.
  • binding of an anti- AD AMTS 13 autoantibody to an ADAMTS13 increases accumulation of VWF or ULVWF.
  • anti- ADAMTS13 autoantibodies or immune complexes comprising the same may reduce and/or inactivate ADAMTS13 thereby causing endothelial cell injury.
  • anti- ADAMTS13 autoantibodies or immune complexes comprising the same may cause platelet activation.
  • binding of an anti- AD AMTS 13 autoantibody to an ADAMTS13 increases microthrombi or platelet-rich clots.
  • a glycoengineered polypeptide disclosed herein or a composition comprising the same binds to one or more autoantibodies, e.g., one or more IgG4 autoantibodies, associated with TTP.
  • an autoantibody associated with TTP is an anti- ADAMTS13 autoantibody (e.g., an IgG4 anti- ADAMTS13 autoantibody).
  • administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same to a subject having or identified with TTP results in a reduction in the level and/or activity of one or more autoantibodies (e.g., one or more IgG4 autoantibodies) in a subject.
  • a reduction in the level and/or activity of one or more autoantibodies is compared to (1) a healthy subject, or (2) the same subject prior to administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same.
  • a glycoengineered polypeptide disclosed herein comprises a first moiety that binds to an IgG4 antibody.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • an anti-ADAMTS13 autoantibody or a fragment or a complex thereof is an anti-ADAMTS13 autoantibody or a fragment or a complex thereof.
  • CIDP Chronic Inflammatory Demyelinating Polyneuropathy
  • Chronic Inflammatory Demyelinating Polyneuropathy is a chronic, progressive immune-mediated inflammatory neuropathy characterized by peripheral nerve demyelination.
  • Typical CIDP is generally associated with symmetrical polyneuropathy which equally affects proximal and distal muscles along with sensory dysfunction.
  • atypical CIDP is generally associated with five different syndromes with separable symptom profiles. These profiles include multifocal acquired demyelinating sensory and motor (MADSAM), distal acquired demyelination symmetric (DADS), pure motor, pure sensory, and focal subtypes (Koike & Katsuno. Neurol. Ther. (2020) 9:213-227).
  • MADSAM multifocal acquired demyelinating sensory and motor
  • DADS distal acquired demyelination symmetric
  • pure motor pure sensory, pure sensory, and focal subtypes
  • CIDP is diagnosed using signs and symptoms as well as electrodiagnostic methods. Therefore, although patients can be assigned a particular subtype based on symptom presentation, patients may convert from one subtype to another, and the underlying pathophysiological mechanisms often overlap (Koike & Katsuno. Neurol. Ther. (2020) 9:213- 227). At present, at least two major mechanisms are believed to underlie demyelination and the resulting neuropathy in CIDP: inflammatory macrophage-mediated demyelination and IgG4- autoantibody-mediated nodo-paranodopathy. Both types of pathophysiology may result in demyelination and impairment of sensory and/or motor function.
  • Efficient transmission of neural signals between motor neurons and muscles and among sensory neurons is critical for healthy motor and sensory ability.
  • Mammalian neurons are covered in myelin, which provides insulation for axons and increases the fidelity of signals between neurons and between neurons and muscles.
  • the clustering of ion channels in nodal and paranodal regions of the axon facilitates active transmission, as they allow the action potential to jump between nodes of Ranvier, further increasing the efficiency of neural communication.
  • neurofascin In healthy neurons, neurofascin, an ankyrin-binding cell adhesion IgG molecule, aids in the growth of neurites, myelination, bundling of neural fibers, and inter-neuronal adhesion (Tait et al. Jour. Cell Bio. (2000) 150(3): 657-666).
  • NF186 In the central nervous system, at least two major isoforms of neurofascin are expressed: NF186 and NF155.
  • the NF186 isoform is typically expressed in neurons, near axon initial segments and nodes of Ranvier, where it interacts with ankyrin-Gto cluster voltage-dependent sodium channels.
  • NF155 is typically expressed in glia, tends to cluster in paranodal regions of the myelinated axonal sheath near axo- glial junctions, and likely mediates interactions between axons and Schwann cells.
  • the IgG molecule contactin-1 interacts with NF155 and contributes to myelination and cellular adhesion at paranodal junctions (Chatterjee et al. Neural Regen. Res. (2019) 14(2): 206-216).
  • IgG4 anti-neurofascin 155 (Anti-NF155) and/or IgG4 anti-contactin 1 antibodies have been observed in paranodal junctions between terminal loops of myelin and near axolemma.
  • anti-neurofascin 140/186 antibodies have been observed in the nodal region.
  • Available treatment options for patients with CIDP include corticosteroids, rituximab, intravenous immunoglobulin, plasma exchange, azathioprine, mycophenolate mofetil, methotrexate, or a combination thereof.
  • none of the available treatment options can reduce the levels of IgG4 autoantibodies or immune complexes comprising the same.
  • Anti-neurofascin 155 (anti-NF155) and/or anti-contactin 1 (anti-CNTNl) antibodies are observed in patients with various subtypes of CIDP.
  • Anti-NF155 and anti-CNTNl antibodies are associated with severely compromised paranodal junctions which typically show increased space between adjacent myelin, increased width of myelin loops, and detachment of terminal loops from axolemma (Lehmann et al. J. Neurol. Neurosurg. Psychiatry (2019) 90:981-987; Koike and Katsuno, Neurol. Ther. (2020) 9:213-227).
  • anti-NF186 antibodies are typically found in the nodal region where they may be associated with impaired clustering of voltage-dependent sodium channels. All three types of autoantibodies (anti-NF155, anti-NF186, and anti-CNTNl) are generally of the IgG4 class.
  • a glycoengineered polypeptide disclosed herein or a composition comprising the same binds to one or more autoantibodies, e.g., one or more IgG4 autoantibodies, associated with CIDP.
  • an autoantibody associated with CIDP is anti- NF155 autoantibody (e.g., an IgG4 anti-NF155 autoantibody).
  • an autoantibody associated with CIDP is anti-NF186 autoantibody (e.g., an IgG4 anti-NF186 autoantibody).
  • an autoantibody associated with CIDP is anti-CNTNl autoantibody (e.g., an IgG4 anti-CNTNl autoantibody).
  • administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same to a subject having or identified with CIDP results in a reduction in the level and/or activity of one or more autoantibodies (e.g., one or more IgG4 autoantibodies) in a subject.
  • a reduction in the level and/or activity of one or more autoantibodies is compared to (1) a healthy subject, or (2) the same subject prior to administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same.
  • a glycoengineered polypeptide disclosed herein comprises a first moiety that binds to an IgG4 antibody.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • an IgG4 antibody is anti-NF186 autoantibody.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • an anti-NF155 autoantibody e.g., IgG4 autoantibody
  • an IgG4 antibody e.g., IgG4 autoantibody
  • MN Membranous Nephropathy
  • Membranous nephropathy is a glomerular disease that can occur at all ages. In adults, it is the most frequent cause of nephrotic syndrome.
  • Membranous nephropathy is an autoimmune disease which is characterized by a thickening of glomerular capillary walls due to immune complex deposition on the subepithelial side of the glomerular basement membrane (GBM).
  • Immune deposits typically comprise of immunoglobulin G (IgG) (e.g., one or more antipodocyte autoantibodies), one or more podocyte autoantigens, and/or one or more complement complexes (e.g., a membrane attack complex [MAC]) (Ronco et al., (2021) “Membranous Nephropathy” Nature Reviews: Disease Primer 7:69).
  • IgG immunoglobulin G
  • MAC membrane attack complex
  • the pathways which can be induced by the complement MAC include: protein kinases, lipid metabolism, reactive oxygen species, growth factors, gene transcription, endoplasmic reticulum stress and the ubiquitin-proteasome system (Ke et al., (2022) BMC Nephrology 23:313).
  • membranous nephropathy can be classified as primary (idiopathic) or secondary membranous nephropathy, which account for 75%-80% and 20%-25% of MN, respectively.
  • Primary or idiopathic membranous (pMN/iMN) nephropathy cases are most common and are often associated with autoantibodies recognizing podocyte autoantigens which can form immune complexes along the glomerular basement membrane (GBM).
  • GBM glomerular basement membrane
  • Secondary membranous nephropathy cases are often associated with autoimmune diseases, malignancies, infections, and/or drugs.
  • a recent report has suggested a role for VEGFA in the pathogenesis of idiopathic membranous nephropathy. According to Ke et al. 2022, VEGFA induced activation of the PI3K-Akt signaling pathway can lead to vascular hyperpermeability resulting in increased filtration of inflammatory factors, complements, and cytokines.
  • Available treatment options for patients with membranous nephropathy include administration of a VEGF pathway inhibitor, an anti-B AFF antibody, an anti-CD20 antibody, an anti-CD38 antibody, an anti-CD19 antibody, a Janus kinase inhibitor, a SYK inhibitor, a Factor B complement inhibitor, or a combination thereof.
  • a VEGF pathway inhibitor an anti-B AFF antibody
  • an anti-CD20 antibody an anti-CD38 antibody
  • an anti-CD19 antibody a Janus kinase inhibitor
  • SYK inhibitor a Factor B complement inhibitor
  • none of the available treatment options reduce and/or remove anti-podocyte autoantibodies or immune complexes comprising the same.
  • Anti-podocyte autoantibodies and podocyte autoantigens include administration of a VEGF pathway inhibitor, an anti-B AFF antibody, an anti-CD20 antibody, an anti-CD38 antibody, an anti-CD19 antibody, a Janus kinase inhibitor, a SYK inhibitor,
  • Membranous nephropathy may be induced by autoimmunity to one or more autoantigens expressed on podocytes (e.g., by the development of anti-podocyte autoantibodies).
  • anti-podocyte autoantibodies or immune complexes comprising the same form deposits at the glomerular basement membrane (GBM).
  • anti-podocyte autoantibodies or immune complexes comprising the same may cause podocyte injury.
  • anti-podocyte autoantibodies or immune complexes comprising the same may cause thickening of the GBM.
  • antipodocyte autoantibodies or immune complexes comprising the same contribute to and/or result in Membranous nephropathy (e.g., idiopathic membranous nephropathy).
  • an anti- podocyte autoantibody is an IgG antibody. In some embodiments, an anti-podocyte autoantibody is an IgG4 antibody. In some embodiments, an anti-podocyte autoantibody does not directly induce complement activation (e.g., complement activation is independent of Fc receptor function). Without wishing to be bound by any particular theory, in some embodiments, where an anti-podocyte autoantibody is an IgG4 autoantibody, complement activation is a result of the formation of immune complexes comprising antipodocyte antibodies and podocyte autoantigens.
  • Podocytes are cells in Bowman's capsule in the kidneys that wrap around capillaries of the glomerulus. Podocytes make up the epithelial lining of Bowman's capsule, the third layer through which filtration of blood takes place.
  • Podocytes express a number of polypeptides (e.g., autoantigens) including but not limited to: phospholipase A2 receptor (PLA2R), Thrombospondin Type-1 Domain-Containing 7A (THSD7A), neutral endopeptidase (NEP), Neural Epidermal Growth Factor like 1 Protein (NELL-1), Exostosin 1 (EXT1), Exostosin 2 (EXT2), Semaphorin 3B (SEMA3B; UniProt/Swiss-Prot Accession No.
  • PHA2R phospholipase A2 receptor
  • TSD7A Thrombospondin Type-1 Domain-Containing 7A
  • NEP neutral endopeptidase
  • NELL-1 Neural Epidermal Growth Factor like 1 Protein
  • EXT1 Exostosin 1
  • EXT2 Exostosin 2
  • SEMA3B Semaphorin 3B
  • NCAM1 Neural Cell Adhesion Molecule 1
  • PCDH7 Protocadherin 7
  • a podocyte autoantigen is expressed in a podocyte, e.g., in the cytoplasm, nucleus, peri-nucleus, or in a compartment in a cell. In some embodiments, a podocyte autoantigen is expressed on the cell surface of podocytes.
  • diseases associated with anti-podocyte autoantibodies such as membranous nephropathy (e.g., idiopathic membranous nephropathy) are associated with increased and/or aberrant expression of one or more podocyte autoantigens.
  • membranous nephropathy e.g., idiopathic membranous nephropathy
  • membranous nephropathy is associated with increased and/or aberrant expression of one or more anti-podocyte autoantibodies or immune complexes comprising the same.
  • a podocyte autoantigen comprises: PLA2R, THSD7A, NEP, NELLI, EXT1, EXT2, SEMA3B, NCAM1, PCDH7, or a combination thereof.
  • a podocyte autoantigen is a PLA2R polypeptide, or a variant or fragment thereof.
  • a podocyte autoantigen is a THSD7A polypeptide, or a variant or fragment thereof.
  • a podocyte autoantigen is a NELLI polypeptide, or a variant or fragment thereof.
  • a podocyte autoantigen is a NEP polypeptide, or a variant or fragment thereof.
  • a podocyte autoantigen is a EXT1 polypeptide, or a variant or fragment thereof.
  • a podocyte autoantigen is a EXT2 polypeptide, or a variant or fragment thereof.
  • a podocyte autoantigen is a SEMA3B polypeptide, or a variant or fragment thereof.
  • a podocyte autoantigen is a NCAM1 polypeptide, or a variant or fragment thereof.
  • a podocyte autoantigen is a PCDH7 polypeptide, or a variant or fragment thereof.
  • a glycoengineered polypeptide disclosed herein or a composition comprising the same binds to one or more autoantibodies, e.g., one or more IgG4 autoantibodies, associated with MN.
  • an autoantibody associated with MN is an anti- PLA2R autoantibody.
  • an autoantibody associated with MN is an anti- THSD7A autoantibody.
  • an autoantibody associated with MN is an anti- NELL autoantibody.
  • an autoantibody associated with MN is an anti-NEP autoantibody.
  • an autoantibody associated with MN is an anti- EXT1 autoantibody.
  • an autoantibody associated with MN is an anti- EXT2 autoantibody.
  • administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same to a subject having or identified with MN results in a reduction in the level and/or activity of one or more autoantibodies (e.g., one or more IgG4 autoantibodies) in a subject.
  • a reduction in the level and/or activity of one or more autoantibodies is compared to (1) a healthy subject, or (2) the same subject prior to administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same.
  • a glycoengineered polypeptide disclosed herein comprises a first moiety that binds to an IgG4 antibody.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • an IgG4 antibody is an anti-PLA2R autoantibody or a fragment or a complex thereof.
  • an IgG4 antibody e.g., IgG4 autoantibody
  • an IgG4 antibody is an anti-NELL autoantibody or a fragment or a complex thereof.
  • an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-NEP autoantibody or a fragment or a complex thereof. In some embodiments, an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-EXTl autoantibody or a fragment or a complex thereof. In some embodiments, an IgG4 antibody (e.g., IgG4 autoantibody) is an anti-EXT2 autoantibody or a fragment or a complex thereof.
  • Immunoglobulin G4-Related Disease IgG4-RD
  • Immunoglobulin G4-Related Disease is a family of chronic, fibroinflammatory disorders (Al-Khalil & Erickson. Missouri Medicine (2016) 115(3): 253-256). IgG4-RD is characterized by the infiltration of IgG4-positive plasma cells into various organs. These infiltrates can cause storiform fibrosis and the development of tumor-like lesions and organ damage. Although IgG4 is also implicated in the pathophysiology of other disorders such as idiopathic membranous glomerulonephritis and pemphigus vulgaris, IgG4-RD is both pathologically and clinically distinct from other diseases (Stone et al. New Eng.
  • IgG4-RD symptoms are heterogenous and depend on the organ and type of tissue involved. For example, IgG4-RD is associated with autoimmune pancreatitis, retroperitoneal fibrosis, and tubulointerstitial nephritis, among numerous other conditions. Many patients develop the disease in multiple organs systems or tissues, either at initial symptom presentation or more often over months or years following initial symptom onset. Across subtypes of IgG4-RD, most patients present with IgG4-positive lesions in the affected organs or tissues as well as allergic disease, such as atopy, eczema, and asthma. In many but not all cases, serum IgG4 is also elevated. IgG4-RD is more likely to affect men and persons over 50 years of age.
  • IgG4-RD In histological analysis, the most distinguishing characteristics of IgG4-RD are typically the presence of dense, IgG4-positive lymphoplasmacytic infiltrates in a storiform pattern, obliterative phlebitis, and eosinophil infiltrates.
  • messenger RNA expression in studies of affected tissue has implicated type 2 helper T (Th2) cells in the initial pathophysiology of IgG4-RD.
  • Th2 cytokines such as interleukin-4, interleukin-5, interleukin- 10, and interleukin- 13 are observed at higher rates in IgG4-RD -affected tissue compared to other autoimmune disorders.
  • Th2 cells can also trigger the activity of regulatory T (Treg) cell, and CD4+CD25+ Treg cells are observed in histological analysis of affected tissue. Furthermore, transforming growth factor-0 is overexpressed in IgG4-RD and may underlie the commonly observed fibrotic processes in the disease (Stone et al. New Eng. Jour. Med., 2012, 366(6): 539- 51).
  • Treg regulatory T
  • IgG4-RD Available treatments for IgG4-RD include glucocorticoids, biliary stenting, methotrexate, mycophenolate mofetil, azathioprine, and rituximab. However, none of the available treatment options specifically target IgG4 antibodies or immune complexes comprising the same.
  • a glycoengineered polypeptide disclosed herein or a composition comprising may be useful in the treatment and/or prevention of IgG4-RD.
  • administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same to a subject having or identified with IgG4-RD results in a reduction in the level and/or activity of IgG4 antibodies in a subject.
  • a reduction in the level and/or activity of IgG4 antibodies is compared to (1) a healthy subject, or (2) a subject prior to administration of a glycoengineered polypeptide disclosed herein or a composition comprising the same.
  • compositions and pharmaceutical compositions are provided.
  • composition disclosed herein may comprise and/or deliver one or more glycoengineered polypeptides disclosed herein or nucleic acids encoding one or more glycoengineered polypeptides disclosed herein.
  • a composition disclosed herein comprises a glycoengineered polypeptide comprising a first moiety and a second moiety. In some embodiments, a composition disclosed herein comprises a plurality of glycoengineered polypeptides comprising a first moiety and a second moiety.
  • a composition comprising a plurality of glycoengineered polypeptides comprises 1, 2, 3, 4, 5, or more glycoengineered polypeptides comprising a first moiety that binds to an IgG4 autoantibody (e.g., a first moiety that binds to the same IgG4 autoantibody).
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti- desmoglein 1 (Dsgl) IgG4 autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-desmoglein 3 (Dsg3) IgG4 autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti- nicotinic acetylcholine receptor (nAChR) IgG4 autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-muscle specific kinase (MuSK) IgG4 autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-low density lipoprotein receptor-related protein 4 (LRP4) IgG4 autoantibody or a fragment thereof.
  • LRP4 anti-low density lipoprotein receptor-related protein 4
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-titin autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-ryanodine receptor autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-agrin receptor autoantibody or a fragment thereof. In some embodiments, a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-collagen Q receptor autoantibody or a fragment thereof. In some embodiments, a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-Kvl.4 potassium channel autoantibody or a fragment thereof. In some embodiments, a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-cortactin autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti- NF155 IgG4 autoantibody or a fragment thereof. In some embodiments, a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-CNTNl IgG4 autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti- PLA2R autoantibody or a fragment thereof. In some embodiments, a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-THSD7A autoantibody or a fragment thereof. In some embodiments, a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-NELL autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-NEP autoantibody or a fragment thereof. In some embodiments, a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-EXTl autoantibody or a fragment thereof. In some embodiments, a composition comprising a plurality of glycoengineered polypeptides comprises glycoengineered polypeptides having a first moiety that binds to an anti-EXT2 autoantibody or a fragment thereof.
  • a composition comprising a plurality of glycoengineered polypeptides comprises 1, 2, 3, 4, 5, or more glycoengineered polypeptides each comprising a first moiety that binds to a different IgG4 antibody (e.g., an IgG4 autoantibody or a fragment thereof).
  • a composition comprising a plurality of glycoengineered polypeptides comprises a first glycoengineered polypeptide comprising a first moiety that binds to an IgG4 autoantibody and a second glycoengineered polypeptide comprising a first moiety that binds to a different IgG4 autoantibody.
  • ratio of the first glycoengineered polypeptide to the second glycoengineered polypeptide is about 1 :9, about 1:8, about 1:7, about 1 :6, about 1:5, about 1 :4, about 1:3, about 1 :2.5, about 1 :2, about 1: 1.5, about 1: 1, about 9:1, about 8: 1, about 7: 1, about 6: 1, about 5:1, about 4: 1, about 3:1, about 2.5:1 about, 2: 1, or about 1.5: 1.
  • ratio of the first glycoengineered polypeptide to the second glycoengineered polypeptide is 1 :9, 1:8, 1 :7, 1:6, 1:5, 1:4, 1:3, 1:2.5, 1:2, 1: 1.5, 1: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3:1, 2.5:1, 2:1, or 1.5: 1.
  • the ratio of the first glycoengineered polypeptide to the second glycoengineered polypeptide is about 1 :5 to about 5: 1; about 1:2.5 to about 2.5:1.
  • the ratio of the first glycoengineered polypeptide to the second glycoengineered polypeptide is about 1 :1.5 to about 1.5: 1.
  • the first glycoengineered polypeptide is present at an amount of about 10-90% and the second glycoengineered polypeptide is present at an amount of about 90- 10%.
  • the first glycoengineered polypeptide is present at an amount of about 20-80% and the second glycoengineered polypeptide is present at an amount of about 80- 20%.
  • the first glycoengineered polypeptide is present at an amount of about 30-70% and the second glycoengineered polypeptide is present at an amount of about 70- 30%.
  • the first glycoengineered polypeptide is present at an amount of about 40-60% and the second glycoengineered polypeptide is present at an amount of about 60- 40%.
  • the first glycoengineered polypeptide is present at an amount of about 10% and the second glycoengineered polypeptide is present at an amount of about 90%.
  • the first glycoengineered polypeptide is present at an amount of about 20% and the second glycoengineered polypeptide is present at an amount of about 80%.
  • the first glycoengineered polypeptide is present at an amount of about 30% and the second glycoengineered polypeptide is present at an amount of about 70%.
  • the first glycoengineered polypeptide is present at an amount of about 40% and the second glycoengineered polypeptide is present at an amount of about 60%.
  • the first glycoengineered polypeptide is present at an amount of about 50% and the second glycoengineered polypeptide is present at an amount of about 50%.
  • the first glycoengineered polypeptide is present at an amount of about 60% and the second glycoengineered polypeptide is present at an amount of about 40%.
  • the first glycoengineered polypeptide is present at an amount of about 70% and the second glycoengineered polypeptide is present at an amount of about 30%. [0622] In some embodiments, the first glycoengineered polypeptide is present at an amount of about 80% and the second glycoengineered polypeptide is present at an amount of about 20%.
  • the first glycoengineered polypeptide is present at an amount of about 90% and the second glycoengineered polypeptide is present at an amount of about 10%.
  • compositions comprising a population of glycoengineered polypeptides, wherein the population of glycoengineered polypeptides has an N-glycan profile that is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or about 100% homogeneous at one or more of the N-glycosylation site(s).
  • the homogeneity of the N-glycan profile at one or more of the N- glycosylation sites is determined by N-glycan analysis, glycopeptide analysis or intact protein analysis.
  • the N-glycan profile comprises about 30% to 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of the N-glycan of the structure provided herein.
  • the population of glycoengineered polypeptides has an N-glycan profile comprising about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% of the N-glycan of the structure provided herein among all glycans in the N-glycan profile.
  • a glycoengineered polypeptide disclosed herein or a composition comprising the same may be useful to treat and/or prevent a disease described herein (e.g., a disease associated IgG4 autoantibodies), or to ameliorate a symptom associated with a disease, disorder or condition described herein.
  • a disease described herein e.g., a disease associated IgG4 autoantibodies
  • ameliorate a symptom associated with a disease, disorder or condition described herein may be useful to treat and/or prevent a disease described herein (e.g., a disease associated IgG4 autoantibodies), or to ameliorate a symptom associated with a disease, disorder or condition described herein.
  • the present disclosure also provides pharmaceutical compositions that, when administered to a subject (e.g., a human subject), e.g., when administered to a subject suffering from a disease associated with IgG4 autoantibodies, deliver a glycoengineered polypeptide as described herein to such subject.
  • a subject e.g., a human subject
  • the present disclosure provides pharmaceutical compositions that comprise or deliver one or more glycoengineered polypeptides or one or more polynucleotides encoding such as described herein.
  • a pharmaceutical composition is or comprises a composition according to the present disclosure.
  • a pharmaceutical composition typically includes a glycoengineered polypeptide as described herein, or a nucleic acid that encodes it in combination with one or more pharmaceutically acceptable carriers or excipients such as, for example one or more buffers, diluents, fillers, salts, solubilizers, stabilizers, and/or other materials as is known in the art.
  • pharmaceutically acceptable carriers or excipients such as, for example one or more buffers, diluents, fillers, salts, solubilizers, stabilizers, and/or other materials as is known in the art.
  • carrier components appropriate to a particular active type (e.g., polypeptide versus nucleic acid, viral vector vs plasmid versus RNA, etc. ⁇ and/or route of administration (e.g., parenteral, enteral, etc. .
  • a pharmaceutical composition may comprise or deliver two or more different glycoengineered polypeptide, so that such agents may be administered in combination (e.g., substantially simultaneously or sequentially) to subject(s).
  • a pharmaceutical composition may contain one or more agents that, for example, may improve stability of the composition and/or its active agent (e.g., to particular storage conditions and/or period(s) of time), facilitate delivery of the composition and/or its active agent, and/or otherwise enhance effectiveness (and/or reduce one or more undesirable side effects) of the active agent or composition once administered.
  • a provided pharmaceutical composition may comprise or deliver another active agent in addition to an glycoengineered polypeptide as described herein.
  • the present disclosure provides methods of treating and/or preventing an IgG4 autoantibody associated disease in a subject comprising administering a glycoengineered polypeptide and/or composition as described herein, thereby improving at least one sign or symptom of the IgG4 autoantibody associated disease in the subject after administration.
  • methods of treating and/or preventing a disease associated with IgG4 autoantibodies comprising administering a glycoengineered polypeptide and/or composition as described herein.
  • a disease associated with IgG4 autoantibodies is an autoimmune disease and/or an inflammatory disorder.
  • an autoimmune disease is pemphigus vulgaris.
  • an autoimmune disease is pemphigus foliaceus.
  • an autoimmune disease is muscle-specific kinase myasthenia gravis.
  • an autoimmune disease is thrombotic thrombocytopenia purpura.
  • an autoimmune disease is chronic inflammatory demyelinating polyneuropathy.
  • an autoimmune disease is membranous nephropathy.
  • an autoimmune disease is primary membranous nephropathy.
  • an autoimmune disease is idiopathic membranous nephropathy.
  • an autoimmune disease is IgG4-Related Disease.
  • glycoengineered polypeptides that specially bind to IgG4 autoantibodies or fragments or complexes thereof.
  • glycoengineered polypeptides disclosed herein have therapeutic value, e.g., in the treatment of a disease associated with IgG4 autoantibodies or fragments or complexes thereof.
  • a glycoengineered polypeptide that specifically binds to an IgG4 autoantibody of the present disclosure can be used, inter alia, to treat, prevent, and/or improve IgG4 autoantibody associated diseases, including but not limited to any number of diseases in which the IgG4 autoantibody levels are aberrantly high and/or in which a reduction of IgG4 autoantibody levels is sought.
  • a subject to be treated with methods described herein can be e.g., a patient having, or at risk of having, or is diagnosed as having a disease associated with IgG4 autoantibodies.
  • a disease associated with IgG4 autoantibodies has or is characterized as having increased levels of IgG4 autoantibodies. In some embodiments, a disease associated with IgG4 autoantibodies has or is characterized as having aberrant IgG4 autoantibodies.
  • a method of treating and/or preventing pemphigus in a subject comprises administering to a subject a glycoengineered polypeptide and/or composition according to the present disclosure.
  • a subject has an anti-Dsgl and/or an anti-Dsg3 autoantibody.
  • administration of a composition reduces a level of anti-Dsgl and/or reduces a level of anti-Dsg3 autoantibody as compared to a subject who has not been administered the composition or as compared to the same subject prior to administration of the composition.
  • a method of treating and/or preventing myasthenia gravis (MG) comprises administering to a subject a glycoengineered polypeptide and/or composition according to the present disclosure.
  • MG myasthenia gravis
  • a subject has an anti-nicotinic acetylcholine receptor (nAchR) IgG4 autoantibody, an anti-muscle specific kinase (MuSK) IgG4 autoantibody, an anti-LRP4 IgG4 autoantibody, an anti-titin autoantibody, an anti-ryanodine receptor IgG4 autoantibody, an anti-agrin IgG4 autoantibody, an anti-collagen Q IgG4 autoantibody, an anti-K v 1.4 potassium channel IgG4 autoantibody and/or an anti-cortactin IgG4 autoantibody.
  • nAchR anti-nicotinic acetylcholine receptor
  • MoSK anti-muscle specific kinase
  • administration of a composition reduces a level of anti-nAchR IgG4 autoantibody, reduces a level of anti-MuSK IgG4 autoantibody, reduces a level of anti-LRP4 IgG4 autoantibody, reduces a level of anti-titin IgG4 autoantibody, reduces a level of anti-ryanodine IgG4 receptor autoantibody, reduces a level of anti-agrin IgG4 autoantibody, reduces a level of anti-collagen Q IgG4 autoantibody, reduces a level of anti- K v 1.4 potassium channel IgG4 autoantibody, and/or reduces a level of anti- cortactin IgG4 autoantibody as compared to a subject who has not been administered the composition or as compared to the same subject prior to administration of the composition.
  • a method of treating and/or preventing thrombotic thrombocytopenic purpura (TTP) (e.g., idiopathic iTTP), in a subject comprises administering to a subject a composition according to the present disclosure.
  • a subject has an anti- ADAMTS13 autoantibody.
  • administration of a composition reduces a level of an anti-ADAMTS13 autoantibody as compared to a subject who has not been administered the composition or as compared to the same subject prior to administration of the composition.
  • a method of treating and/or preventing chronic inflammatory demyelinating polyneuropathy (CIDP) comprises administering to a subject a glycoengineered polypeptide and/or composition according to the present disclosure.
  • a subject has an anti-NF155 and/or an anti-CNTNl autoantibody.
  • administration of a composition reduces a level of an anti-NF155 autoantibody and/or reduces a level of an anti-CNTNl autoantibody as compared to a subject who has not been administered the composition or as compared to the same subject prior to administration of the composition.
  • a method of treating and/or preventing membranous neuropathy (MN) comprises administering to a subject a glycoengineered polypeptide and/or composition according to the present disclosure.
  • MN membranous neuropathy
  • a subject has an anti-PLA2R autoantibody, an anti-THSD7A autoantibody, an anti-NELL autoantibody, an anti -NEP autoantibody, an anti -EXT 1 autoantibody, or an anti-EXT2 autoantibody.
  • administration of a composition reduces a level of an anti-PLA2R autoantibody, reduces a level of an anti-THSD7A autoantibody, reduces a level of an anti-NELL autoantibody, reduces a level of an anti-NEP autoantibody, reduces a level of an anti-EXTl autoantibody and/or reduces a level of an anti- EXT2 autoantibody as compared to a subject who has not been administered the composition or as compared to the same subject prior to administration of the composition.
  • a method of treating and/or preventing IgG4-Related Disease in a subject comprises administering to a subject a glycoengineered polypeptide and/or composition according to the present disclosure.
  • a subject has an IgG4 autoantibody.
  • administration of a composition reduces a level of an IgG4 autoantibody as compared to a subject who has not been administered the composition or as compared to the same subject prior to administration of the composition.
  • a glycoengineered polypeptide and/or composition according to the present invention is used to reduce and/or degrade IgG4 autoantibodies.
  • a glycoengineered polypeptide and/or composition according to the present invention is used to reduce the risk of a disease associated with IgG4 autoantibodies.
  • a glycoengineered polypeptide and/or composition according to the present invention is used to ameliorate one or more symptoms of a disease associated with IgG4 autoantibodies.
  • a method comprising, assessing a level of an IgG4 autoantibody in a sample from a subject, and administering a glycoengineered polypeptide and/or pharmaceutical composition disclosed herein, if the level of the IgG4 autoantibody is higher than a comparator.
  • a comparator comprises a predetermined reference sample such as a sample obtained from an otherwise similar subject who does not have a disease or disorder, or a symptom of a disease or disorder.
  • an IgG4 autoantibody comprises: an anti-desmoglein 1 (Dsgl) autoantibody or a fragment or a complex thereof; or an anti-desmoglein 3 (Dsg3) autoantibody or a fragment or a complex thereof.
  • Dsgl anti-desmoglein 1
  • Dsg3 anti-desmoglein 3
  • an IgG4 autoantibody comprises: an anti-nicotinic acetylcholine receptor (nAchR) autoantibody or a fragment or a complex thereof; an anti-muscle-specific kinase (MuSK) IgG4 autoantibody or a fragment or a complex thereof; an anti-low-density lipoprotein receptor-related protein 4 (LRP4) autoantibody or a fragment or a complex thereof; an anti-titin autoantibody or a fragment or a complex thereof; an anti-ryanodine receptor autoantibody or a fragment or a complex thereof; an anti-agrin autoantibody or a fragment or a complex thereof; an anti-collagen Q autoantibody or a fragment or a complex thereof; an anti- K V 1.4 potassium channel autoantibody or a fragment or a complex thereof; or an anti-cortactin autoantibody or a fragment or a complex thereof.
  • nAchR anti-nicotinic acetylcholine receptor
  • an IgG4 autoantibody comprises: an anti- AD AMTS 13 autoantibody or a fragment or a complex thereof.
  • an IgG4 autoantibody comprises: an anti-neurofascin 155 (NF 155) autoantibody or a fragment or a complex thereof; or an anti-contactin 1 (CNTN1) autoantibody or a fragment or a complex thereof.
  • NF 155 anti-neurofascin 155
  • CNTN1 anti-contactin 1
  • an IgG4 autoantibody comprises: an anti-PLA2R autoantibody or a fragment or a complex thereof; anti-THSD7A autoantibody or a fragment or a complex thereof; an anti-NELL autoantibody or a fragment or a complex thereof; an anti-NEP autoantibody or a fragment or a complex thereof; an anti-EXTl autoantibody or a fragment or a complex thereof; or an anti-EXT2 autoantibody or a fragment or a complex thereof.
  • a method comprises administering to a subject a pharmaceutical composition comprising a glycoengineered polypeptide according to the present disclosure.
  • Delivery of a glycoengineered polypeptide can be achieved e.g., by administration of a pharmaceutical composition as described herein, such as a pharmaceutical composition that comprises a glycoengineered polypeptide or a nucleic acid that encodes it, for example via oral ingestion, inhalation, topical application or parenteral administration (e.g., cutaneous, subcutaneous, intraperitoneal, intramuscular or intravenous injection).
  • parenteral administration e.g., cutaneous, subcutaneous, intraperitoneal, intramuscular or intravenous injection.
  • administration is by intravenous or intramuscular injection.
  • local administration may be or comprise topical administration (e.g., to the skin) or parenteral administration (e.g., by injection to a site of deposition such as to the kidney).
  • delivery of a glycoengineered polypeptide can be achieved e.g., by administration of a pharmaceutical composition as described herein, such as a pharmaceutical composition that comprises a glycoengineered polypeptide or a nucleic acid that encodes it, may be oral, rectal, ophthalmic (including intravitreal or intracameral), nasal, topical (including buccal and sublingual), intrauterine, vaginal or parenteral (including subcutaneous, intraperitoneal, intramuscular, intravenous, intradermal, intracranial, intratracheal, and epidural).
  • ophthalmic including intravitreal or intracameral
  • nasal including buccal and sublingual
  • intrauterine vaginal or parenteral
  • parenteral including subcutaneous, intraperitoneal, intramuscular, intravenous, intradermal, intracranial, intratracheal, and epidural.
  • compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.
  • the administration step comprises intravenous injection, intraperitoneal injection, subcutaneous injection, transdermal injection, or intramuscular injection.
  • composition according to the present disclosure are delivered to a subject suffering from or susceptible to a disease associated with IgG4 antibodies as described herein.
  • the subject has or is diagnosed as having pemphigus vulgaris (PV) or pemphigus foliaceus (PF).
  • the subject has or is diagnosed as having myasthenia gravis, e.g., muscle-specific kinase myasthenia gravis (MuSK-MG).
  • the subject has or is diagnosed as having thrombotic thrombocytopenic purpura (TTP), e.g., idiopathic TTP (iTTP).
  • TTP thrombotic thrombocytopenic purpura
  • iTTP idiopathic TTP
  • the subject has or is diagnosed as having chronic inflammatory demyelinating polyneuropathy (CIDP), e.g. typical CIDP or atypical CIDP.
  • CIDP chronic inflammatory demyelinating polyneuropathy
  • the subject has or is diagnosed as having membranous neuropathy (MN), e.g., idiopathic MN (iMN) or primary MN (pMN).
  • MN membranous neuropathy
  • iMN idiopathic MN
  • pMN primary MN
  • IgG4-RD IgG4-Related Disease
  • the individual is a human.
  • a subject has or is diagnosed as having IgG4 autoantibodies .
  • a subject having IgG4 autoantibodies is treated with a glycoengineered polypeptide or a nucleic acid that encodes it that specifically binds IgG4 autoantibodies.
  • administration of a composition according to the present disclosure alleviates one or more symptoms of pemphigus vulgaris (PV) or pemphigus foliaceus (PF).
  • administration of a composition according to the present disclosure alleviates one or more symptoms of myasthenia gravis, e.g., muscle-specific kinase myasthenia gravis (MuSK/MG).
  • administration of a composition according to the present disclosure alleviates one or more symptoms of thrombotic thrombocytopenic purpura (TTP), e.g., idiopathic TTP (iTTP).
  • TTP thrombotic thrombocytopenic purpura
  • administration of a composition according to the present disclosure alleviates one or more symptoms of chronic inflammatory demyelinating polyneuropathy (CIDP), e.g., typical CIDP or atypical CIDP.
  • CIDP chronic inflammatory demyelinating polyneuropathy
  • administration of a composition according to the present disclosure alleviates one or more symptoms of membranous neuropathy (MN), e.g., idiopathic MN (iMN) or primary MN (pMN).
  • MN membranous neuropathy
  • iMN idiopathic MN
  • pMN primary MN
  • administration of a composition according to the present disclosure alleviates one or more symptoms of IgG4-Related Disease (IgG4-RD).
  • IgG4-RD IgG4-Related Disease
  • administration of a glycoengineered polypeptide that binds to an anti-Dsgl autoantibody or a fragment or a complex thereof treats and/or prevents pemphigus foliaceus.
  • administration of a glycoengineered polypeptide that binds to an anti-Dsg3 autoantibody treats and/or prevents pemphigus vulgaris.
  • administration of a glycoengineered polypeptide that binds to an anti-nicotinic acetylcholine receptor (nAchR) autoantibody treats and/or prevents myasthenia gravis, e.g. MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti-muscle specific kinase IgG4 autoantibody or a fragment or a complex thereof treats and/or prevents myasthenia gravis, e.g., MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti-low-density lipoprotein receptor-related protein 4 (LRP4) autoantibody or a fragment or a complex thereof treats and/or prevents myasthenia gravis, e.g., MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti-titin autoantibody or a fragment or a complex thereof treats and/or prevents myasthenia gravis, e.g., MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti-ryanodine receptor autoantibody or a fragment or a complex thereof treats and/or prevents myasthenia gravis, e.g., MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti-agrin autoantibody or a fragment or a complex thereof treats and/or prevents myasthenia gravis, e.g., MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti-collagen Q autoantibody or a fragment or a complex thereof treats and/or prevents myasthenia gravis, e.g., MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti-K v 1.4 potassium channel autoantibody or a fragment or a complex thereof treats and/or prevents myasthenia gravis, e.g., MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti- cortactin autoantibody or a fragment or a complex thereof treats and/or prevents myasthenia gravis, e.g., MuSK-MG.
  • administration of a glycoengineered polypeptide that binds to an anti-ADAMTS13 autoantibody or a fragment or a complex thereof treats and/or prevents thrombotic thrombocytopenia purpura (TTP), e.g., idiopathic TTP (iTTP).
  • TTP thrombotic thrombocytopenia purpura
  • iTTP idiopathic TTP
  • administration of a glycoengineered polypeptide that binds to an anti-neurofascin 155 (NF 155) autoantibody or a fragment or a complex thereof treats and/or prevents chronic inflammatory neuropathy (CIDP).
  • administration of a glycoengineered polypeptide that binds to an anti-contactin 1 (CNTN1) autoantibody or a fragment or a complex thereof treats and/or prevents thrombotic thrombocytopenia purpura (TTP), e.g., idiopathic TTP (iTTP).
  • TTP thrombotic thrombocytopenia purpura
  • administration of a glycoengineered polypeptide that binds to an anti-podocyte autoantibody or a fragment or a complex thereof treats and/or prevents membranous nephropathy (MN), e.g., idiopathic MN or primary MN.
  • MN membranous nephropathy
  • administration of a glycoengineered polypeptide that binds to an anti-PLA2R autoantibody or a fragment or a complex thereof treats and/or prevents membranous nephropathy (MN), e.g., idiopathic MN or primary MN.
  • administration of a glycoengineered polypeptide that binds to an anti-THSD7A autoantibody or a fragment or a complex thereof treats and/or prevents membranous nephropathy (MN), e.g., idiopathic MN or primary MN.
  • MN membranous nephropathy
  • administration of a glycoengineered polypeptide that binds to an anti-NELL autoantibody or a fragment or a complex thereof treats and/or prevents membranous nephropathy (MN), e.g., idiopathic MN or primary MN.
  • administration of a glycoengineered polypeptide that binds to an anti-NEP autoantibody or a fragment or a complex thereof treats and/or prevents membranous nephropathy (MN), e.g., idiopathic MN or primary MN.
  • MN membranous nephropathy
  • administration of a glycoengineered polypeptide that binds to an anti-EXTl autoantibody or a fragment or a complex thereof treats and/or prevents membranous nephropathy (MN), e.g., idiopathic MN or primary MN.
  • administration of a glycoengineered polypeptide that binds to an anti-EXT2 autoantibody or a fragment or a complex thereof treats and/or prevents membranous nephropathy (MN), e.g., idiopathic MN or primary MN.
  • MN membranous nephropathy
  • administration of a glycoengineered polypeptide that binds to an IgG4 autoantibody or a fragment or a complex thereof treats and/or prevents IgG4-Related Disease.
  • a method comprises administering a composition once. In some embodiments, a method comprises administering a composition repeatedly (e.g., two or more times over a period of time).
  • administration of a composition is continued to maintain remission (e.g., keep IgG4 autoantibodies low and/or undetectable) and/or avoid relapse.
  • Amounts of glycoengineered polypeptide administered in a single dose may depend on the nature and/or severity of the condition being treated and/or on the nature of prior treatments that the patient has undergone. In some embodiments, the attending physician decides the amount of glycoengineered polypeptide with which to treat each individual patient. In some embodiments, the attending physician initially administers low doses of glycoengineered polypeptides of the present invention and observe the patient's response. In some embodiments, larger doses are administered until an optimal therapeutic effect is obtained for the patient, after which dosage is not increased further.
  • a glycoengineered polypeptide according to the present disclosure is delivered in an amount effective to reduce levels of IgG4 autoantibody or a fragment or a complex thereof.
  • glycoengineered polypeptides may be administered in combination with one or more therapies (e.g., a pharmaceutical agent, dialysis, etc).
  • glycoengineered polypeptides may be administered in combination with one or more pharmaceutical agents.
  • a glycoengineered polypeptide may be administered in combination with one or more therapeutic agents for IgG4 autoantibody associated diseases (such as agents that ameliorate symptoms of IgG4 autoantibody associated diseases), and/or in combination with one or more other pharmaceutical agents.
  • glycol- engineered polypeptides may be administered in combination with one or more therapies and/or agents that are prescribed by a clinician for treatment of IgG4 autoantibody associated diseases.
  • the present disclosure provides, among other things, glycoengineered polypeptides that specifically bind to-IgG4 autoantibodies and thereby causes degradation of IgG4 autoantibodies.
  • IgG4 autoantibodies degradation comprises internalized into a cell for degradation (e.g., by transporting the IgG4 autoantibody to a lysosome).
  • glycoengineered polypeptides specifically binding IgG4 autoantibodies according to the present disclosure are characterized in that they inhibit the biological function of IgG4 autoantibodies (e.g., binding to IgG4).
  • a glycoengineered polypeptide according to the present disclosure is used to degrade and/or reduce the levels of IgG4 autoantibodies. In some embodiments, a glycoengineered polypeptide is used to lower IgG4 autoantibody levels, such as lowering elevated plasma IgG4 autoantibody levels in a subject with a disease associated with IgG4 autoantibodies as described herein.
  • the present disclosure provides glycoengineered polypeptides characterized in that when administered to a cell, tissue, or subject, the glycoengineered polypeptide which is bound to a target via the first moiety and to an endocytic receptor via a second moiety results in degradation of the target.
  • degradation comprises internalization into a cell. In some embodiments, degradation comprises lysosomal degradation. In some embodiments, degradation occurs in a liver cell.
  • the present disclosure provides glycoengineered polypeptides characterized in that when administered to a cell, tissue, or subject, a glycoengineered polypeptide which is bound to a target via the first moiety and to an endocytic receptor via a second moiety is targeted (e.g., distributed) to a liver cell and/or tissue.
  • a glycoengineered polypeptide which is bound to a target via the first moiety and to an endocytic receptor via a second moiety is targeted (e.g., distributed) to a liver cell and/or tissue.
  • an endocytic receptor bound by a second moiety is or comprises ASGPR.
  • ASGPR can be detected on the surface of a liver cell or tissue and/or in a liver cell or tissue.
  • targeting results in degradation of a glycoengineered polypeptide and a complex comprising the same (e.g., a complex comprising a glycoengineered polypeptide and a target) in a liver cell and/or tissue.
  • degradation comprises lysosomal degradation.
  • the present disclosure provides glycoengineered polypeptides characterized in that when administered to a cell, tissue, or subject, a glycoengineered polypeptide which is bound to a target via the first moiety and to an endocytic receptor via a second moiety is distributed at lower levels to non-liver cells and/or tissue as compared to distribution of glycoengineered polypeptides to liver cells and/or tissue.
  • a lower level comprises at least 1.5-fold, at least 2-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least 100-fold, or at least 500-fold lower.
  • a non-liver cell and/or tissue comprises spleen cells or tissue, heart cells or tissue, kidney cells or tissue, lung cells or tissue, etc.
  • distribution of glycoengineered polypeptides in one or more cells and/or tissue can be assessed using methods known in the field including methods disclosed herein, e.g., as provided in Example 16.
  • the present disclosure provides glycoengineered polypeptides characterized in that when administered to a cell, tissue, or subject, a glycoengineered polypeptide which is bound to a target via the first moiety and to an endocytic receptor via a second moiety is detected in a liver cell and/or tissue.
  • a glycoengineered polypeptide cannot be readily detected or is detected at lower levels in a non-liver cell and/or tissue.
  • a lower level comprises at least 1.5-fold, at least 2-fold, at least 10- fold, at least 20-fold, at least 50-fold, at least 100-fold, or at least 500-fold lower.
  • a non-liver cell and/or tissue comprises spleen cells or tissue, heart cells or tissue, kidney cells or tissue, lung cells or tissue, etc.
  • the present disclosure provides glycoengineered polypeptides characterized in that when administered to a cell, tissue, or subject, the glycoengineered polypeptide which is bound to a target via the first moiety prevents binding of the target to an immune cell.
  • a target is an IgG4 autoantibody or a fragment thereof.
  • a target comprises an IgG4 autoantibody or a fragment thereof.
  • glycoengineered polypeptides disclosed herein are characterized in that when administered to a cell, tissue or subject, glycoengineered polypeptides or compositions comprising the same reduce the level and/or activity of an autoantibody or a fragment thereof (e.g., an IgG4 autoantibody or a fragment thereof).
  • the reduction is as compared to a comparable cell, tissue or subject administered a different therapeutic agent or not administered any therapeutic agent.
  • the reduction is as compared to the same cell, tissue or subject prior to administration of a glycoengineered polypeptides or compositions comprising the same.
  • a comparator is the same cell, tissue or subject prior to administration of a glycoengineered polypeptide or a composition comprising the same.
  • a comparator is an otherwise comparable cell, tissue or subject who has not been administered the glycoengineered polypeptide or a composition comprising the same.
  • a comparator is an otherwise similar cell, tissue or subject who has been administered a different composition, e.g., different glycoengineered polypeptide or different therapeutic composition, e.g., standard of care.
  • administration of a glycoengineered polypeptide disclosed herein results in a reduction of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99%, in a level and/or activity of an IgG4 autoantibody or a fragment or complex thereof relative to a comparator.
  • administration of a glycoengineered polypeptide disclosed herein results in a reduction of about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100% of a level and/or activity of an IgG4 autoantibody or a fragment or complex thereof relative to a comparator.
  • administration of a glycoengineered polypeptide disclosed herein results in a reduction of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of a level and/or activity of an IgG4 autoantibody or a fragment or complex thereof relative to a comparator.
  • administration of a glycoengineered polypeptide disclosed herein results in a reduction of about 5% to about 100%, about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, or about 90% to about 100%, about 5% to about 100%, about 5% to about 90%, about 5% to about 80%, about 5% to about 70%, about 5% to about 60%, about 5% to about 50%, about 5% to about 40%, about 5% to about 30%, about 5% to about 20%, or about 5% to about 10% of a level and/or activity of an IgG4 autoantibody or a fragment or complex thereof relative to a comparator.
  • a first moiety comprising an antibody agent disclosed herein binds to an IgG4 antibody at an affinity of about lOpM to about lOOOnM, about lOpM to about 500nM, about lOpM to about 400nM, about lOpM to about 300nM, about lOpM to about 200nM, about lOpM to about lOOnM, about lOpM to about lOnM, about lOpM to about InM, about lOpM to about 500pM, about lOpM to about 400pM, about lOpM to about 300pM, about lOpM to about 200 pM, about lOpM to about lOOpM, lOpM to about 50pM, about 50pM to about lOOOnM, about lOOpM to about lOOOnM, about 200pM to about lOOOnM, about 300pM to about
  • a first moiety comprising an antibody agent disclosed herein binds to an IgG4 antibody at an affinity of about 50pM, about lOOpM, about 200pM, about 300pM, about 400pM, about 500pM, about 600pM, about 700pM, about 800pM, about 900pM, about InM, about lOnM, about 20nM, about 50nM, about lOOnM, about 200nM, about 300nM, or about 500nM.
  • a first moiety comprising an antibody agent disclosed herein binds to an IgG4 antibody at an affinity of 50pM, lOOpM, 200pM, 300pM, 400pM, 500pM, 600pM, 700pM, 800pM, 900pM, InM, lOnM, 20nM, 50nM, lOOnM, 200nM, 300nM, or 500nM.
  • a first moiety comprising an anti-IgG4 antibody agent disclosed herein binds to an IgG4 antibody with an affinity of less than 0.2nM. In some embodiments, a first moiety comprising an anti-IgG4 antibody agent disclosed herein binds to an IgG4 antibody with an affinity of about 50pM to about 200pM, about 60pM to about 200pM, about 70pM to about 200pM, about 80pM to about 200pM, about 90pM to about 200pM, about 1 OOpM to about 200pM, about 150pM to about 200pM, about 50pM to about 150pM, about 50pM to about lOOpM, about 50pM to about 90pM, about 50pM to about 80pM, about 50pM to about 70pM, about 50pM to about 60pM.
  • a first moiety comprising an anti-IgG4 antibody agent disclosed herein with a binding affinity to IgG4 of less than 0.2nM is a Bin-1 antibody agent.
  • a Bin-1 antibody agent is clone IG15, clone IG47, clone IG5, clone IG33, clone IG9, clone IG40, clone IG11, clone IG37, clone IG13, clone IG35, clone IG16, clone IG31, clone IG21, clone IG10, or clone IG25.
  • a first moiety comprising an anti-IgG4 antibody agent disclosed herein binds to an IgG4 antibody with an affinity of about 0.20 InM to about 7nM, about 0.4nM to about 7nM , about 0.8nM to about 7nM , about InM to about 7nM , about 2nM to about 7nM , about 3nM to about 7nM , about 4nM to about 7nM , about 5nM to about 7nM .
  • a first moiety comprising an anti-IgG4 antibody agent disclosed herein with a binding affinity to IgG4 of about 0.201nM to about 7nM is a Bin-2 antibody.
  • a Bin-2 antibody agent is clone IG2, clone IG46, clone IG4, clone IG44, clone IG34, clone IG42, clone IG19, clone IG38, clone IG1, clone IG48, clone IG36, clone IG18, clone IG45, clone IG12, clone IG30, clone IG3, clone IG23, clone IG26, clone IG22, clone IG27, or clone IG28.
  • a first moiety comprising an anti-IgG4 antibody agent disclosed herein binds to an IgG4 antibody with an affinity of more than 7nM.
  • a first moiety comprising an anti-IgG4 antibody agent disclosed herein with a binding affinity to IgG4 of more than 7nM is a Bin-3 binder.
  • a Bin-3 binder is a clone IG24, clone IG6, clone IG43, clone IG7, clone IG41, clone IG29, clone IG39, clone IG14, clone IG8, clone IG17, clone IG32, or clone IG20.
  • a first moiety comprising an antibody agent disclosed herein binds to an IgG4 antibody with an affinity of at least 2-fold higher, at least 5-fold higher, at least 10- fold higher, at least 15-fold higher, at least 20-fold higher, at least 30-fold higher, at least 40-fold higher, at least 50-fold higher, at least 60-fold higher, at least 80-fold higher, at least 100-fold higher, at least 150-fold higher, at least 200-fold higher, at least 250-fold higher, at least 300-fold higher, at least 350-fold higher, at least 400-fold higher, at least 450-fold higher, at least 500-fold higher, at least 600-fold higher, at least 700-fold higher, at least 800-fold higher, at least 900-fold higher, at least 1,000-fold higher, at least 2,000-fold higher, at least 3,000-fold higher, at least 4,000-fold higher, at least 5,000-fold higher, at least 6,000-fold higher, at least 7,000-fold higher, at least 8,000-fold
  • a first moiety comprising an antibody agent disclosed herein binds to an IgG4 antibody with an affinity of about 2-fold to about 10,000-fold, about 5-fold to about 10,000-fold, about 10-fold to about 10,000-fold, about 100-fold to about 10,000-fold, about 200- fold to about 10,000-fold, about 500-fold to about 10,000-fold, about 1000-fold to about 10,000-fold, about 2000-fold to about 10,000-fold, about 5000-fold to about 10,000-fold, about 2-fold to about 5,000-fold, about 2-fold to about 2000-fold, about 2-fold to about 1000-fold, about 2-fold to about 500-fold, about 2-fold to about 200-fold, about 2-fold to about 100-fold, about 2-fold to about 50-fold, about 2-fold to about 20-fold, about 2-fold to about 10-fold compared to binding affinity of the first moiety to a non-IgG4 antibody when detected by surface plasmon resonance (SPR).
  • SPR surface plasmon resonance
  • a first moiety comprising an antibody agent disclosed herein binds to an IgG4 antibody with an affinity of about 2-fold to about 100,000-fold, compared to binding affinity of the first moiety to a non-IgG4 antibody when detected by surface plasmon resonance (SPR).
  • SPR surface plasmon resonance
  • the present disclosure provides methods of making a glycoengineered polypeptide comprising a first moiety comprising one or more peptides that specifically binds to a target (e.g., an IgG4 autoantibody or a fragment or a complex thereof) and a second moiety comprising one or more glycans conjugated to the first moiety.
  • a target e.g., an IgG4 autoantibody or a fragment or a complex thereof
  • a glycoengineered polypeptide disclosed herein can be made using methods disclosed in U.S. Provisional Patent Application No. 63/410,955 filed on September 28, 2022, U.S.
  • Section 5.3 discloses Leishmania host cells
  • Section 5.4 discloses exemplary methods of genetically engineering a Leishmania cell for expressing glycoengineered polypeptides
  • Section 5.5 discloses exemplary methods of culturing Leishmania host cells
  • Section 5.6 discloses exemplary uses of Leishmania host cells as an expression system.
  • Section 7.1 discloses Leishmania host cells including modifications that can be made to a Leishmania host cell for producing glycoengineered polypeptides
  • Section 7.2 disclose methods of genetically engineering Leishmania host cells for producing glycoengineered polypeptides
  • Section 7.3 discloses methods of culturing Leishmania host cells.
  • An exemplary method of making glycoengineered polypeptides using Leishmania host cells is provided in Example 1 herein.
  • exemplary Leishmania strains that can be used to make glycoengineered polypeptides disclosed herein include: StCGP3558, StCGP4564, StCGP5359, or StCGP5942.
  • StCGP3558 As would be understood by persons with ordinary skill in the art, such methods and host cells can also be used for making glycoengineered polypeptides disclosed herein.
  • the one or more glycans of the second moiety are conjugated to the first moiety at one or more glycosylation sites with in vivo glycosylation, e.g., in a cell.
  • a cell is a Leishmania host cell.
  • a cell is a glycoengineered yeast host cell, e.g., glycoengineered Pichia pastoris host cell.
  • the one or more glycans of the second moiety are conjugated to the first moiety at one or more glycosylation sites with chemical conjugation, e.g., using Click chemistry.
  • a method for producing a glycoengineered polypeptide comprising (i) culturing a Leishmania host cell under conditions suitable for polypeptide production and (ii) isolating said glycoengineered polypeptide.
  • the Leishmania host cell comprises: (a) a recombinant nucleic acid encoding a glycoengineered polypeptide; and (b) a recombinant nucleic acid encoding one or more recombinant N-acetylgalactosamine (GalNAc) transferases.
  • the Leishmania host cell is capable of producing glycoengineered polypeptide comprising a biantennary, GalNAc-terminated N-glycan.
  • the Leishmania host cells provided herein is capable of producing glycoengineered polypeptide comprising an N-glycan of the following structure: wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N-acetylglucosamine (GlcNAc) residue and the black circle represents a mannose (Man) residue, and wherein X represents an amino acid residue of the glycoengineered polypeptide.
  • the glycoengineered polypeptide produced by the Leishmania host cell is a therapeutic polypeptide, e.g., a polypeptide used in the treatment of a disease or disorder.
  • the glycoengineered polypeptide produced by the Leishmania host cell can be peptide or an antibody.
  • Leishmania host cells for the production of glycoengineered polypeptides disclosed herein, or a population of glycoengineered polypeptides, wherein the Leishmania host cells comprise: (a) a recombinant nucleic acid encoding a glycoengineered polypeptide disclosed herein; and (b) a recombinant nucleic acid encoding one or more recombinant N-acetylgalactosamine (GalNAc) transferases.
  • the Leishmania host cells provided herein are capable of producing glycoengineered polypeptides comprising a biantennary, GalN Ac-terminated N-glycan.
  • the Leishmania host cells provided herein are capable of producing glycoengineered polypeptides comprising an N-glycan of the following structure: wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N-acetylglucosamine (GlcNAc) residue and the black circle represents a mannose (Man) residue, and wherein X represents an amino acid residue of the glycoengineered polypeptide.
  • the Leishmania host cells provided herein comprise a recombinant nucleic acid encoding one or more recombinant N-acetylgalactosamine (GalNAc) transferases disclosed herein.
  • the Leishmania host cells provided herein comprise a recombinant nucleic acid encoding one or more additional recombinant glycosyltransferases disclosed herein.
  • one or more endogenous enzymes disclosed herein from the glycan biosynthesis pathway of the the Leishmania host cells provided herein have been deleted, mutated and/or functionally inactivated.
  • the Leishmania host cells provided herein further comprise a recombinant nucleic acid encoding heterologous UDP-GalNAc biosynthetic pathway proteins capable of generating UDP-GalNAc.
  • the Leishmania host cells provided herein comprise a recombinant nucleic acid encoding a heterologous UDP-GalNAc transporter protein capable of transporting UDP-GalNAc to the secretory pathway.
  • the Leishmania host cells provided herein have been genetically engineered such that the formation of an O-linked GlcNAc on a polypeptide produced in the Leishmania host cell is reduced or eliminated.
  • Leishmania host cells that have been genetically engineered to reduce or eliminate the formation of an O-linked GlcNAc are described, for example, in WO 2021/140143, which is incorporated herein by reference in its entirety.
  • the Leishmania host cells provided herein below are genetically engineered using the methods described herein. In certain embodiments, the Leishmania host cells provided herein below are cultured according to the methods described herein.
  • Suitable host cells comprise liver cells, myeloid cells, immune cells, endothelial cells, parenchymal cells or epithelial cells.
  • the immune cell is a dendritic cell, a macrophage, a monocyte, a microglia cell, a granulocyte or a B lymphocyte.
  • the Leishmania host cells are cultured using any of the standard culturing techniques known in the art. For example, cells are routinely grown in rich media like Brain Heart Infusion, Trypticase Soy Broth or Yeast Extract, all containing 5 pg /ml Hemin. Additionally, incubation is done at 26°C in the dark as static or shaking cultures for 2-3 days. In some embodiments, cultures of recombinant cell lines contain the appropriate selective agents. Non-limiting exemplary selective agents are provided in Table 3.
  • Table 3 Selective agents used during transfection (50% concentration for preselection and 100% concentration for main selection) and standard culturing of L. tarentolae. Double amounts of the selective agents could be used if higher selection pressure was intended.
  • the Leishmania host cells are cultured in a growth medium comprising GalNAc.
  • the growth medium comprises at least 1 mM, at least 2 mM, at least 3 mM, at least 4 mM, at least 5 mM, at least 6 mM, at least 7 mM, at least 8 mM, at least 9 mM, at least 10 mM, at least 11 mM, at least 12 mM, at least 13 mM, at least 14 mM, at least 15 mM, at least 16 mM, at least 17 mM, at least 18 mM, at least 19 mM, or at least 20 mM GalNAc.
  • the growth medium comprises about 1 mM to about 5 mM, about 5 mM to about 10 mM, about 10 mM to about 15 mM, or about 15 mM to about 20 mM GalNAc. In certain embodiments, the growth medium comprises about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, or about 20 mM GalNAc.
  • the growth medium comprises about about 10 mM GalNAc.
  • the Leishmania host cells are cultured in a growth medium comprising GlcNAc.
  • the growth medium comprises at least 1 mM, at least 2 mM, at least 3 mM, at least 4 mM, at least 5 mM, at least 6 mM, at least 7 mM, at least 8 mM, at least 9 mM, at least 10 mM, at least 11 mM, at least 12 mM, at least 13 mM, at least 14 mM, at least 15 mM, at least 16 mM, at least 17 mM, at least 18 mM, at least 19 mM, or at least 20 mM GlcNAc.
  • the growth medium comprises about 1 mM to about 5 mM, about 5 mM to about 10 mM, about 10 mM to about 15 mM, or about 15 mM to about 20 mM GlcNAc. In certain embodiments, the growth medium comprises about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, or about 20 mM GlcNAc.
  • Leishmania host cell may be used as an expression system for making a glycoengineered polypeptide disclosed herein or a population of glycoengineered polypeptides.
  • the glycoengineered polypeptide degrader may be a heterologous, non-Leishmania protein, such as a therapeutic protein (e.g., an antibody).
  • a therapeutic protein e.g., an antibody
  • Other methods of producing Leishmania host cells for use as expression systems are known and may also be used, for example, see WO 2019/002512, WO 2021/140144 and WO 2021/140143, each of which are incorporated herein by reference in their entirety.
  • Use of Leishmania host cells to make monoclonal antibodies are also known. Exemplary methods are described in WO 2022/053673, which is incorporated herein by reference in its entirety.
  • compositions comprising the Leishmania host cells can comprise additional components suitable for maintenance and survival of the Leishmania host cells, and can additionally comprise additional components required or beneficial to the production of glycoengineered bifunctional degraders by fas Leishmania host cells, e.g., inducers for inducible promoters, such as arabinose, IPTG.
  • inducers for inducible promoters such as arabinose, IPTG.
  • yeast or filamentous fungal host cells for the production of glycoengineered polypeptides disclosed herein, or a population of glycoengineered polypeptides.
  • a yeast or filamentous fungal host cell is a K. lactis host cells.
  • a yeast or filamentous fungal host cell is a Pichia pastoris host cell.
  • a yeast or filamentous fungal host cell is a Pichia methanolica host cell.
  • a yeast or filamentous fungal host cell is a Hansenula host cell.
  • Exemplary yeast or filamentous fungal host cells that can be used to produce glycoengineered polypeptides disclosed herein are disclosed in U.S. Patent 8,206,949, the entire contents of which are hereby incorporated by reference.
  • Exemplary yeast or filamentous fungal host cells that can be used to produce glycoengineered polypeptides disclosed herein are disclosed in U.S. Patent7,981,660, the entire contents of which are hereby incorporated by reference.
  • Exemplary yeast or filamentous fungal host cells that can be used to produce glycoengineered polypeptides disclosed herein are disclosed in U.S. Patent 8,883,483, the entire contents of which are hereby incorporated by reference.
  • a yeast or filamentous fungal host cell has been genetically modified to produce glycoproteins with a predominant N-glycan glycoform.
  • the yeast or filamentous fungal host cells provided herein are capable of producing glycoengineered polypeptides comprising a biantennary, GalNAc- terminated N-glycan.
  • the yeast or filamentous fungal host cells provided herein are capable of producing glycoengineered polypeptides comprising an N-glycan of the following structure: wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N-acetylglucosamine (GlcNAc) residue and the black circle represents a mannose (Man) residue, and wherein X represents an amino acid residue of the glycoengineered polypeptide.
  • Embodiment 1 A glycoengineered polypeptide comprising: (a) a first moiety comprising one or more peptides that specifically binds to a human immunoglobulin G4 (IgG4) antibody; and (b) a second moiety comprising one or more glycans conjugated to the first moiety at one or more glycosylation sites.
  • IgG4 immunoglobulin G4
  • Embodiment 2 The glycoengineered polypeptide of embodiment 1, wherein the IgG4 antibody is an IgG4 autoantibody or a fragment or a complex thereof.
  • Embodiment 3 The glycoengineered polypeptide of embodiment 1 or 2, wherein the first moiety comprises one or more peptides that binds the IgG4 antibody at a CHI domain, a hinge domain, a CH2 domain, a CH3 domain, or any combination thereof.
  • Embodiment 4 The glycoengineered polypeptide of embodiment 3, wherein the first moiety binds the IgG4 antibody at a CH2 domain and/or a CH3 domain.
  • Embodiment 5 The glycoengineered polypeptide of any one of embodiments 1-4, wherein the first moiety comprises one or more peptides that binds the IgG4 antibody with a higher affinity as compared to binding to a non-IgG4 antibody.
  • Embodiment 6 The glycoengineered polypeptide of embodiment 5, wherein the first moiety binds to an IgG4 autoantibody with an affinity that is at least 10-fold stronger as compared to binding of the first moiety to a non-IgG4 antibody.
  • Embodiment 7 The glycoengineered polypeptide of embodiment 5 or 6, wherein the non- IgG4 antibody comprises an antibody of a class other than IgG4 (e.g., an IgGl antibody, an IgG2 antibody, an IgG3 antibody, an IgA antibody, an IgM antibody, an IgD antibody, or an IgE antibody, or a fragment or complex of any of the foregoing).
  • a class other than IgG4 e.g., an IgGl antibody, an IgG2 antibody, an IgG3 antibody, an IgA antibody, an IgM antibody, an IgD antibody, or an IgE antibody, or a fragment or complex of any of the foregoing.
  • Embodiment 8 The glycoengineered polypeptide of any one of embodiment 5-7, wherein the first moiety substantially does not bind to a non-IgG4 antibody.
  • Embodiment 9. The glycoengineered polypeptide of any one of embodiments 1-8, wherein the first moiety binds to an IgG4 antibody with an affinity of about lOpM to about lOOOnM.
  • Embodiment 10 The glycoengineered polypeptide of any one of embodiments 1-9, wherein the first moiety binds to an IgG4 antibody with an affinity of at least 2-fold higher compared to binding affinity of the first moiety to a non-IgG4 antibody when detected by surface plasmon resonance (SPR).
  • SPR surface plasmon resonance
  • Embodiment 11 The glycoengineered polypeptide of any one of the preceding embodiments, characterized in that administration of the glycoengineered polypeptide to a cell, tissue, or subject reduces the level of the IgG4 antibody as compared to: (1) an otherwise similar cell, tissue, or subject that has not been administered the glycoengineered polypeptide; or (2) the same cell, tissue or subject, prior to administration of the glycoengineered polypeptide.
  • Embodiment 12 The glycoengineered polypeptide of embodiment 11, wherein the reduction in IgG4 antibody is at least 5%.
  • Embodiment 13 The glycoengineered polypeptide of embodiment 11 , wherein the reduction in IgG4 antibody is about 10% to about 99%.
  • Embodiment 14 The glycoengineered polypeptide of any one of embodiments 5-13, characterized in that administration of the glycoengineered polypeptide to a cell, tissue, or subject does not alter (e.g., does not reduce) the level of a non-IgG4 antibody as compared to: (1) an otherwise similar cell, tissue, or subject that has not been administered the glycoengineered polypeptide; or (2) the same cell, tissue or subject, prior to administration of the glycoengineered polypeptide.
  • Embodiment 15 The glycoengineered polypeptide of embodiment 14, wherein at least 75% of a non-IgG4 antibody remains (e.g., is detectable) after administration of the glycoengineered polypeptide as compared to a level measured prior to the administration.
  • Embodiment 16 The glycoengineered polypeptide of any one of the preceding embodiments, wherein the first moiety comprises an antibody agent.
  • Embodiment 17 The glycoengineered polypeptide of embodiment 16, wherein the antibody agent comprises an antigen binding fragment.
  • Embodiment 18 The glycoengineered polypeptide of embodiment 16 or 17 wherein the antibody agent comprises a full antibody, a Fab fragment, an scFv, a nanobody, a duobody, or a single domain antibody (e.g., a VHH).
  • Embodiment 19 The glycoengineered polypeptide of embodiment 18, wherein the antibody agent is or comprises a Fab fragment.
  • Embodiment 20 The glycoengineered polypeptide of any one of embodiments 16-19, wherein the antibody agent of the first moiety comprises one, two and/or three light chain complementarity determining regions (LC CDRs) and/or one, two and/or three heavy chain complementary determining regions (HC CDRs).
  • LC CDRs light chain complementarity determining regions
  • HC CDRs heavy chain complementary determining regions
  • Embodiment 21 The glycoengineered polypeptide of embodiment 20, wherein the antibody agent of the first moiety comprises a LC CDR1, a LC CDR2 and a LC CDR3.
  • Embodiment 22 The glycoengineered polypeptide of embodiment 21, comprising: (i) a LC CDR1, LC CDR2, and LC CDR3 sequence provided in Table 1; (ii) a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to an LC CDR1, LC CDR2, and LC CDR3 sequence provided in Table 1; or (iii) a sequence having at least 5 substitutions relative to an LC CDR1 , LC CDR2, and LC CDR3 sequence provided in Table 1.
  • Embodiment 23 The glycoengineered polypeptide of any one of embodiments 20-22, wherein the antibody agent of the first moiety comprises a HC CDR1, a HC CDR2 and a HC CDR3.
  • Embodiment 24 The glycoengineered polypeptide of embodiment 23, comprising: (i) a HC CDR1, HC CDR2, and HC CDR3 sequence provided in Table 2; (ii) a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to an HC CDR1, HC CDR2, and HC CDR3 sequence provided in Table 2; or (iii) a sequence having at least 5 substitutions relative to an HC CDR1, HC CDR2, and HC CDR3 sequence provided in Table 2.
  • Embodiment 25 The glycoengineered polypeptide of any one of embodiments 16-24, wherein the antibody agent of the first moiety comprises: (a) a light chain (LC) comprising a sequence provided in Table 1, or a sequence with at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity thereto or a sequence having at least 5, 10, or 20 substitutions relative to a VL region provided in Table 1, optionally wherein the LC polypeptide further comprises one or more framework regions, and/or a constant region, further optionally wherein the light chain constant region comprises a lambda light chain constant region or a variant or portion thereof; or a kappa light chain constant region or a variant or a portion thereof; and/or (b) a heavy chain (HC) comprising a
  • Embodiment 26 The glycoengineered polypeptide of any one of embodiments 2-25, wherein the IgG4 autoantibody comprises: (i) an anti-PLA2R IgG4 autoantibody or a fragment or a complex thereof; (ii) an anti-THSD7A IgG4 autoantibody or a fragment or a complex thereof; (iii) an anti-NELL IgG4 autoantibody or a fragment or a complex thereof; (iv) an antiNEP IgG4 autoantibody or a fragment or a complex thereof; (v) an anti-EXTl IgG4 autoantibody or a fragment or a complex thereof; and/or (vi) an anti-EXT2 IgG4 autoantibody or a fragment or a complex thereof.
  • the IgG4 autoantibody comprises: (i) an anti-PLA2R IgG4 autoantibody or a fragment or a complex thereof; (ii) an anti-THSD7A IgG4 autoantibody
  • Embodiment 27 The glycoengineered polypeptide of any one of embodiments 2-25, wherein the IgG4 autoantibody comprises an anti- AD AMTS 13 IgG4 autoantibody or a fragment or a complex thereof.
  • Embodiment 28 The glycoengineered polypeptide of any one of embodiments 2-25, wherein the IgG4 autoantibody comprises an anti-desmoglein 1 IgG4 autoantibody or a fragment or a complex thereof; and/or an anti-desmoglein 3 IgG4 autoantibody or a fragment or a complex thereof.
  • Embodiment 29 The glycoengineered polypeptide of any one of embodiments 2-25, wherein the IgG4 autoantibody comprises: (i) an anti-nicotinic acetylcholine receptor (nAChR) IgG4 autoantibody or a fragment or a complex thereof;(ii) an anti-muscle-specific kinase (MuSK) IgG4 autoantibody or a fragment or a complex thereof; and/or (iii) an anti-low-density lipoprotein receptor-related protein 4 (LRP4) IgG4 autoantibody or a fragment or a complex thereof.
  • nAChR anti-nicotinic acetylcholine receptor
  • MoSK anti-muscle-specific kinase
  • LRP4 anti-low-density lipoprotein receptor-related protein 4
  • Embodiment 30 The glycoengineered polypeptide of any one of the preceding embodiments, wherein the second moiety comprising one or more glycans specifically binds to one or more endocytic receptors.
  • Embodiment 31 The glycoengineered polypeptide of embodiment 30, wherein the endocytic receptor is or comprises an endocytic lectin.
  • Embodiment 32 The glycoengineered polypeptide of embodiment 31, wherein the endocytic receptor is chosen from: an asialoglycoprotein receptor (ASGPR); a mannose binding receptor, a Cluster of Differentiation 206 (CD206) receptor; a DC-SIGN (Cluster of Differentiation 209 or CD209) receptor; a C-Type Lectin Domain Family 4 Member G (LSECTin) receptor; a macrophage inducible Ca2+-dependent lectin receptor (Mincle); a L- SIGN CD209L receptor; dectin- 1; dectin -2, langerin, macrophage mannose 2 receptor, BDCA- 2, DCIR, MBL, MDL, MICE, CLEC2, DNGR1, CLEC12B, DEC-205, and mannose 6 phosphate receptor (M6PR), or a combination thereof.
  • ASGPR asialoglycoprotein receptor
  • CD206 Cluster of Differentiation 206
  • DC-SIGN Cluster of Differentiation 209
  • Embodiment 33 The glycoengineered polypeptide of any one of the preceding embodiments wherein the one or more glycans comprises a terminal GlcNac.
  • Embodiment 34 The glycoengineered polypeptide of any one of the preceding embodiments, wherein the one or more glycans comprises a terminal GalNac.
  • Embodiment 35 The glycoengineered polypeptide of any one of the preceding embodiments, wherein the one or more glycans comprises a terminal Gal.
  • Embodiment 36 The glycoengineered polypeptide of any one of the preceding embodiments, wherein the one or more glycans is an N-glycan.
  • Embodiment 37 The glycoengineered polypeptide of embodiment 36, wherein the N- glycan is linked to the glycoengineered polypeptide at 1, 2, 3, 4 or 5 N-glycosylation sites.
  • Embodiment 38 The glycoengineered polypeptide of any one of the preceding embodiments, wherein the one or more glycans comprise a glycan structure comprising GlcNAc2-Man3 -GlcNAc2, GalNAc2-GlcNAc2-Man3 -GlcNAc2, Gal2-GlcNAc2-Man3 - GlcNAc2, GlcNAcl-Man3-GlcNAc2, Gal2-GlcNAc2-Man3-GlcNAc2, Gall - GlcNAc2-Man3- GlcNAc2, GalNAcl-GlcNAc2-Man3-GlcNAc2, GlcNAc3-Man3-GlcNAc2, GlcNAc4-Man3- GlcNAc2, Gal3-GlcNAc3-Man3-GlcNAc2, GalNAc3-GlcNAc3-Man3-GlcNAc2, GalNAc4- GlcNAc2, Gal3-
  • Embodiment 39 The glycoengineered polypeptide of embodiment 38, wherein increasing a number of glycan structures on the glycoengineered polypeptide increases the rate of lysosomal degradation as compared to an otherwise similar glycoengineered polypeptide with fewer glycan structures.
  • Embodiment 40 The glycoengineered polypeptide of embodiment 38, wherein a number of glycan structures comprise 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more or 10 or more.
  • Embodiment 41 The glycoengineered polypeptide of any one of embodiments 38-40, wherein the glycan structure comprises a monoantennary structure, biantennary structure, a triantennary structure, or a tetraantennary structure.
  • Embodiment 42 The glycoengineered polypeptide of embodiment 41, wherein the glycan structure comprises a biantennary structure,
  • Embodiment 43 The glycoengineered polypeptide of embodiment 41, wherein the glycan structure comprises a biantennary GalNAc.
  • Embodiment 44 The glycoengineered polypeptide of embodiment 43, wherein the biantennary GalNac binds to an asialoglycoprotein receptor (ASGPR) or a fragment or variant thereof, or a complex comprising ASGPR.
  • ASGPR asialoglycoprotein receptor
  • Embodiment 45 The glycoengineered polypeptide of any one of embodiments 36-44, wherein the N-glycan has a structure of: wherein the black square represents an N-acetyl galactosamine (GalNAc), the white square represents an N-acetylglucosamine (GlcNAc) residue and the black circle represents a mannose (Man) residue, and wherein X represents an amino acid residue of the first moiety.
  • GalNAc N-acetyl galactosamine
  • Man mannose
  • Embodiment 46 The glycoengineered polypeptide of any one of embodiments 36-45, wherein the N-glycan is conjugated to the glycoengineered polypeptide at at least one, two, three, or four N-glycosylation sites.
  • Embodiment 47 The glycoengineered polypeptide of any one of embodiments 36-46, wherein the N-glycan is conjugated to the glycoengineered polypeptide at one, two, three, or four N-glycosylation sites.
  • Embodiment 48 The glycoengineered polypeptide of embodiment 46 or 47, wherein the N-glycosylation site comprises a consensus sequence of N-X-S/T or N-X-C, wherein X is any amino acid except proline.
  • Embodiment 49 The glycoengineered polypeptide of any one of embodiments 46-48, wherein the N-glycosylation site is naturally occurring.

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Abstract

La présente invention concerne des polypeptides glycomodifiés comprenant une première fraction comprenant un ou plusieurs peptides qui se lient de manière spécifique à un anticorps anti-IgG4 (par exemple un auto-anticorps anti-IgG4) et une seconde fraction comprenant un ou plusieurs glycanes conjugués à la première fraction au niveau d'un ou de plusieurs sites de glycosylation. L'invention concerne en outre des compositions comprenant des polypeptides glycomodifiés et/ou des acides nucléiques codant pour ceux-ci, ainsi que leurs procédés de fabrication et d'utilisation.
PCT/IB2025/053675 2024-04-08 2025-04-08 Polypeptides glycomodifiés ciblant des auto-anticorps anti-igg4 et leurs utilisations Pending WO2025215516A1 (fr)

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