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WO2018070942A1 - Formation directe de motifs en dessous de 20 nm de protéine recombinante par fabrication de transfert de motif - Google Patents

Formation directe de motifs en dessous de 20 nm de protéine recombinante par fabrication de transfert de motif Download PDF

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Publication number
WO2018070942A1
WO2018070942A1 PCT/SG2017/050516 SG2017050516W WO2018070942A1 WO 2018070942 A1 WO2018070942 A1 WO 2018070942A1 SG 2017050516 W SG2017050516 W SG 2017050516W WO 2018070942 A1 WO2018070942 A1 WO 2018070942A1
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WIPO (PCT)
Prior art keywords
protein
film
thick
template
suckerin
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PCT/SG2017/050516
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English (en)
Inventor
Shawn HOON
Zhaogang DONG
Joel Kwang Wei Yang
Rubayn GOH
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Agency for Science Technology and Research Singapore
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Agency for Science Technology and Research Singapore
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00—Manufacture or treatment of nanostructures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/735—Fusion polypeptide containing domain for protein-protein interaction containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)

Definitions

  • the present invention generally relates to nano technology.
  • the present invention relates to nanostructures and manufacture of nanostructures.
  • Biopolymers are increasingly attractive as green alternatives for constructing biocompatible and biodegradable electronic/photonic devices. These biopolymers have direct applicability for biomedical applications, such as implantable devices.
  • One approach is to use materials that integrate both the structural and functional optical components into a single element. This application places certain requirements on the substrate material, such as biocompatibility, transparency, toughness, flexibility, and others.
  • a method of manufacturing a protein film comprising a patterned nanostructure, wherein the protein that is forming the protein film is a self-assembling protein comprising: (a) applying a protein solution to a template shaped to correspond with the patterned nanostructure, wherein the template comprises structures with a width of 20 nm and below; (b) allowing the proteins in the protein solution to self-assemble to form a protein film; and (c) removing the protein film from the template.
  • a method of manufacturing a protein-metal hybrid film comprising a patterned nanostructure, wherein the protein that is forming the protein film is a self-assembling protein comprising: (a) applying a protein solution to a template shaped to correspond with the patterned nanostructure, wherein the template comprises structures with a width of 20 nm and below; (b) allowing the proteins in the protein solution to self-assemble to form a protein film; (c) removing the protein from the template; and (d) applying a metal layer to a surface of the protein film, wherein the surface comprises the patterned nanostructure.
  • the protein is a suckerin protein.
  • the suckerin protein is selected from the group consisting of a suckerin monomer, a suckerin trimer, a suckerin pentamer, and a suckerin octamer.
  • the template comprises structures with a width of more than Onm to ⁇ 20nm.
  • the structures are protrusions.
  • the structures are of regular shape or wherein the structures are of regular shape selected from the group consisting of triangle, square, rectangle, and hexagon.
  • the structures in the template are spaced apart from each other at a distance from about lOnm to about 400nm.
  • the protein solution comprises from about 0.5 wt% to about 5 wt% of protein.
  • the template is a template for an optical device.
  • the template is made of silicon.
  • (b) is carried out in vacuum.
  • (b) comprises allowing the proteins in the protein solution to self-assemble for more than 0 hour to up to 24 hours.
  • the protein film is of about ⁇ to about 500 ⁇ thick.
  • removing the protein film from the template in (c) comprises lifting the protein film with mechanical tools or allowing the protein film to attach to an adhesive material which is brought in contact with the protein film and removed after attachment of the protein film to the adhesive material.
  • the protein-metal hybrid film is a protein-gold hybrid film.
  • (d) comprises applying a gold layer to the surface of the protein film.
  • applying a gold layer comprises evaporating gold films to the surface of the protein film.
  • the gold films are of about lnm to about 300nm thick.
  • the present invention provides a protein film comprising a patterned nano structure, wherein the protein is a self-assembling protein, and the patterned nano structure has a width of 20nm and below.
  • the protein film is manufactured according to the method as described herein.
  • the present invention provides a protein-metal hybrid film comprising a patterned nanostructure, wherein the protein is a self-assembling protein, and the patterned nanostructure has a width of 20nm and below.
  • the protein-metal hybrid film is manufactured according to the method as described herein.
  • the nanostructure is biocompatible and/or biodegradable.
  • the protein film as described herein or the protein-metal hybrid film as described herein further comprising a polymer or an active agent.
  • the present invention provides an optical device comprising a protein-metal hybrid film as defined herein.
  • Figure 1 shows data related to genetic engineering of Suckerins.
  • Figure 1A shows a set of amino acid sequences depicting the molecular architecture of two suckerins from Dosidicus gigas (DG-19) and Sepioteuthis lessoniana (SL-lb). The amino acid residues of beta-sheet forming regions are bolded. The amino acid residues of amorphous glycine rich regions are italicized. Proline residues that flank the beta-sheet forming regions are underlined.
  • Figure IB shows a graphical illustration depicting schematic of multimeric suckerins generated by genetic engineering.
  • Figure 1C shows a Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) result depicting the molecular weight analysis of multimeric suckerins produced by bacterial production.
  • Figure 1 illustrates examples of the generated suckerin multimers.
  • Figure 2 shows a schematic illustration depicting one example of the fabrication process of hybrid protein-gold nanostructures.
  • Figure 2A shows a graphical illustration depicting a silicon template with sub-20-nm-wide protrusions.
  • the silicon templates were fabricated by using inductively-coupled plasma (ICP) drying etching based on the mask of hydrogen silsesquioxane (HSQ) resist defined by electron beam lithography (EBL).
  • Figure 2B shows a graphical illustration depicting suckerin protein film as casted onto the sub-20- nm silicon template.
  • Figure 2C shows a graphical illustration depicting the tape placed onto the suckerin protein resist and followed by the template stripping process.
  • Figure 2D shows a graphical illustration depicting the patterned suckerin protein nanostructures with sub-20-nm gaps.
  • Figure 2E shows a graphical illustration depicting the evaporation of gold film onto the suckerin protein nanostructures.
  • Figures 2A to 2D illustrate an exemplary method of manufacturing a protein film comprising a patterned nanostructure
  • Figures 2A to 2E illustrate an exemplary method of manufacturing a protein-gold hybrid film comprising a patterned nanostructure.
  • Figure 3 shows a set of Scanning Electron Micrograph (SEM) images depicting suckerin protein resist having sub-20-nm dimensions of different shapes.
  • Figure 3A shows a Scanning Electron Micrograph (SEM) image depicting a silicon template generated by electron beam lithography.
  • Figures 3B, 3C, and 3D show a set of Scanning Electron Micrograph (SEM) images depicting free-standing suckerin protein film casted onto silicon templates having square (Figure 3B), triangular (Figure 3C) and hexagonal (Figure 3D) features.
  • Figure 3 illustrates that the patterned protein nanostructures can have various shapes whilst maintaining gap size of less than 20 nm.
  • Figure 4 shows data depicting optical characterization results of the hybrid protein-gold nanostructures.
  • Figure 4A shows an optical microscope image depicting hybrid protein-gold nanostructures having varying pitch and varying dosage. The color and the color intensity of each square vary. As the dose time increases and the pitch size ( ⁇ ) decreases, the color and the color intensity becomes darker. The reflectance optical spectrum of each square
  • Figure 4B shows a set of spectra depicting the reflectance optical spectrum of features (i.e. the squares in Figure 4A) with varying pitch size (from 60nm to 360nm; the difference of the pitch size between one reflectance optical spectrum to the next is 20nm) outlined by dotted box in Figure 4A.
  • the dashed line labeled as "Dip.” on Figure 4B marked the position of the observable dips in the optical reflectance spectrum. The dip position refers to the wavelength of having minimum reflectance value.
  • FIG. 4 shows a Scanning Electron Micrograph (SEM) image of Feature 9 (i.e. a feature having a dose time of 2.5 ⁇ 8 and a pitch size of 220 nm, which correspond to the ninth square from the bottom right corner of Figure 4A, inside of the dotted box) depicting the evidence of the presence of sub-20-nm gap size in the features even after the gold evaporation process.
  • SEM Scanning Electron Micrograph
  • Figure 5 shows a schematic illustration depicting one example of a representative manufacturing process of hybrid protein-metal nanostructures.
  • Figure 5A shows a graphical illustration depicting a template with sub-20-nm-wide protrusions.
  • Figure 5B shows a graphical illustration depicting protein film as casted onto the sub-20-nm template.
  • Figure 5C shows a graphical illustration depicting the adhesive material placed onto the protein resist and followed by the template stripping process.
  • Figure 5D shows a graphical illustration depicting the patterned protein nanostructures with sub-20-nm gaps.
  • Figure 5E shows a graphical illustration depicting the application of metal film onto the protein nanostructures.
  • Figures 5A to 5D illustrate an exemplary method of manufacturing a protein film comprising a patterned nanostructure
  • Figures 5A to 5E illustrate an exemplary method of manufacturing a protein-metal hybrid film comprising a patterned nanostructure.
  • Figure 6 shows a schematic illustration depicting one example of cross section of a portion of a template and a portion of a corresponding protein film that is manufactured using said template.
  • the size of the gap (or trenches) of the protein film corresponds to the size of the structures on template.
  • Figure 6A shows a portion a template and
  • Figure 6B shows a portion of a protein film.
  • the schematic illustration is not drawn to scale.
  • Figure 7 shows a set of spectra depicting the result of molecular weight determination of monomeric, trimeric, pentameric, and octameric forms of the suckerin protein using Matrix Assisted Laser Desorption/Ionization - Time Of Flight (MALDI-TOF) analysis.
  • Figure 7 A shows a MALDI-TOF spectrum corresponding to the monomeric form of suckerin that has molecular weight of about 14.9KDa.
  • Figure 7B shows a MALDI-TOF spectrum corresponding to the trimeric form of suckerin that has molecular weight of about 37.7KDa.
  • Figure 7C shows a MALDI-TOF spectrum corresponding to the pentameric form of suckerin that has molecular weight of about 60.4KDa.
  • Figure 7D shows a MALDI-TOF spectrum corresponding to the octameric form of suckerin that has molecular weight of about 93.8KDa.
  • Figure 7 illustrates that even though the repetitive nature of the proteins causes them to migrate differently during SDS-PAGE (as shown in Figure 1C), MALDI- TOF verifies the predicted molecular weight of the proteins.
  • Biopolymers such as proteins are becoming increasingly attractive as green alternatives for constructing biocompatible and biodegradable electronic and photonic devices, which have direct applicability to biomedical applications, such as implantable devices.
  • one approach is to use materials that integrate both the structural and functional optical components into a single element. This design places certain requirements on the substrate materials, such as biocompatibility, transparency, toughness, flexibility, and the like.
  • biocompatibility e.g., biocompatibility
  • transparency e.g., polyethylene
  • toughness e.g., polystymers
  • the inventors of the present disclosure have found an alternative method of manufacturing a protein film comprising a patterned nanostructure and a protein-metal hybrid film comprising a patterned nanostructure.
  • the present disclosure describes the direct patterning of protein (such as genetically engineered suckerin) into nanostructures with dimension of sub-20-nm, and the realization of compact optical color filters by protein- metal (such as protein-gold) hybrid nanostructures.
  • protein-metal hybrid film such as protein-gold
  • the second variant of the method can be thought of as an inverse of the first variant.
  • the protein film is obtained and it is then imprinted (or stamped or impressed) with a template.
  • the step of removing the protein film from the template is identical for both the first variant and the second variant of the method of manufacturing.
  • a protein film comprising a patterned nanostructure is obtained.
  • the third variant of the manufacturing method comprises providing a protein-film manufactured according to the first variant followed by the step of applying a metal layer on the patterned surface of the protein-film.
  • the fourth variant of the manufacturing method comprises providing a protein-film manufactured according to the second variant followed by the step of applying a metal layer on the patterned surface of the protein-film.
  • the present disclosure provides a method of manufacturing a protein film comprising a patterned nanostructure, wherein the protein that is forming the protein film is a self-assembling protein, the method comprising: (a) applying a protein solution to a template shaped to correspond with the patterned nanostructure, wherein the template comprises structures with a width of 20 nm and below; (b) allowing the proteins in the protein solution to self-assemble to form a protein film; and (c) removing the protein film from the template.
  • the present disclosure provides a method of manufacturing a protein film comprising a patterned nanostructure, wherein the protein that is forming the protein film is a self-assembling protein, the method comprising: (a) obtaining a protein film comprising the self-assembling protein; (b) imprinting the protein film from (a) with a template shaped to correspond with the patterned nanostructure to form a patterned nanostructure on the protein film, wherein the template comprises structures with a width of 20 nm and below; and (c) removing the protein film from the template.
  • the present disclosure provides method of manufacturing a protein-metal hybrid film comprising a patterned nanostructure, wherein the protein that is forming the protein film is a self-assembling protein, the method comprising: (a) applying a protein solution to a template shaped to correspond with the patterned nanostructure, wherein the template comprises structures with a width of 20 nm and below; (b) allowing the proteins in the protein solution to self-assemble to form a protein film; (c) removing the protein from the template; and (d) applying a metal layer to a surface of the protein film, wherein the surface comprises the patterned nanostructure.
  • the present disclosure provides a method of manufacturing a protein-metal hybrid film comprising a patterned nanostructure, wherein the protein that is forming the protein film is a self-assembling protein, the method comprising: (a) obtaining a protein film comprising the self-assembling protein; (b) imprinting the protein film from (a) with a template shaped to correspond with the patterned nanostructure to form a patterned nanostructure on the protein film, wherein the template comprises structures with a width of 20 nm and below; and (c) removing the protein film from the template; and (d) applying a metal layer to a surface of the protein film, wherein the surface comprises the patterned nanostructure.
  • applying a protein solution refers to any suitable method for putting or spreading a protein solution on the patterned surface of a template.
  • “applying a protein solution” is performed by dripping the protein solution using an instrument such as a pipette and allowing the protein solution to dry on the template.
  • the proteins that are applicable for manufacturing biocompatible and biodegradable electronic and photonic devices such as a protein film and/or a protein metal hybrid film described herein, have to possess certain characteristics.
  • the self-assembling protein is capable of forming non-covalent nano-confined beta sheets.
  • self-assembling protein refers to a protein that can aggregate or self- aggregates in a protein solution.
  • the protein itself will form the nanostructures on the surface of the protein-film by conforming to the template.
  • the intrinsic property of a protein to form non-covalent nano-confined beta sheets is useful for the formation of free-standing films that can be molded in to sub-20nm features.
  • the self-assembling protein includes, but is not limited to, a silk protein, a suckerin protein, and the like.
  • the silk protein can originate from any source. In one example, the silk protein originates from silkworm or spider or the like.
  • the protein used in the method of manufacturing described herein is a suckerin protein.
  • Suckerins are one family of proteins recently identified in the teeth as found in the suckers that line the arm and tentacles of squids (or the sucker teeth of squids). These teeth play an important grappling role in squid predation.
  • Suckerins self-assemble into robust supramolecular networks via hydrogen bonding and thus can be used to produce materials that have mechanical properties capable of matching with synthetic polymers, despite being composed solely of proteins held together by weak non-covalent interactions.
  • these proteins can be produced recombinantly and easily processed to form functional components such as hydrogels and films with a range of mechanical properties through controlled cross-linking.
  • suckerins are molecularly organized in a hierarchical manner, comprising repeating units of silk- like beta sheet forming motifs and amorphous glycine -rich domains.
  • the inventors have surprisingly found that modifying the numbers of repeating units (i.e. the number of monomers) that a suckerin protein has can affect the toughness and the flexibility of the protein film or the protein-metal hybrid film obtained from the suckerin protein.
  • the suckerin protein includes, but is not limited to, a suckerin monomer, a suckerin dimer, a suckerin trimer, a suckerin tetramer, a suckerin pentamer, a suckerin hexamer, a suckerin heptamer, a suckerin octamer, a suckerin nonamer, and the like.
  • a suckerin monomer, a suckerin trimer, a suckerin pentamer, and a suckerin octamer have been synthesized.
  • the suckerin proteins described herein are exemplified in Figure 1C.
  • suckerin proteins used for the method of manufacturing described herein have been obtained via genetic modification.
  • the suckerin dimer, or the suckerin trimer, or the suckerin tetramer, or the suckerin pentamer, or the suckerin hexamer, or the suckerin heptamer, or the suckerin octamer, or the suckerin nonamer, or the like is obtained by genetic modification of the wild type suckerin monomer.
  • the wild type suckerin can originate from any organism that produces suckerin proteins, such as squids. It is commonly known in the art that there is common species of squid that give rise to almost all known squid species.
  • the wild type suckerin monomer is from an organism (such as a squid) that includes, but is not limited to, Dosidicus gigas (DG-19), Sepioteuthis lessoniana (SL-lb), Sepia esculenta, Loligo vulgaris, Loligo pealei, Todarodes pacificus, Euprymna scolopes, and the like.
  • DG-19 Dosidicus gigas
  • SL-lb Sepioteuthis lessoniana
  • Sepia esculenta Loligo vulgaris
  • Loligo pealei Loligo pealei
  • Todarodes pacificus Euprymna scolopes, and the like.
  • the inventors have surprisingly found that in order to obtain localized lateral gap plasmon resonance or to achieve a strong localized plasmon resonance, a protein-metal hybrid film manufactured according to the method described herein needs to have sub-20nm gap size.
  • the term "localized plasmon resonance” refers to the excitation of the collective oscillation of electrons on metal.
  • the localized plasmon resonance can be measured using any method known in the art. In one example, the localized plasmon resonance is measured by using the micro -spectrometer to measure the reflectance spectra. The localized plasmon resonances lie on the wavelength position, which is corresponding to the dip wavelength in the reflectance spectrum.
  • the optical reflectance spectra of the samples can also be measured using any method known in the art.
  • the optical reflectance spectra of the samples were measured by using a CRAIC UV-VIS-NIR micro- spectrophotometer model QDI 2010 (equipped with a 36x objective lens with NA of 0.5).
  • the size of the gaps on the surface of a protein-film or a protein-metal hybrid film depends of the width of the structures on the template used in the manufacturing process described herein.
  • the template comprises structures with a width of about 20nm, or about 19nm, or about 18nm, or about 17nm, or about 16nm, or about 15nm, or about 14nm, or about 13nm, or about 12nm, or about l lnm, or about lOnm, or about 9nm, or about 8nm, or about 7nm, or about 6nm, or about 5nm, or about 4nm, or about 3nm, or about 2nm, or about lnm, or below about 20nm, or below about 19nm, or below about 18nm, or below about 17nm, or below about 16nm, or below about 15nm, or below about 14nm, or below about 13nm, or below about 12nm, or below about l lnm, or below about lOnm, or below about 9nm, or below about 8nm, or below about 7nm, or below about 6nm, or below about 5nm,
  • the structures comprised on a template used in the manufacturing method described herein are protrusions.
  • the term "protrusions on a template” refers to structures that are extending from the surface of a template. Due to their sizes, the structures that are extending from the surface of a template (or the protrusions on a template) are considered as nanostructures.
  • the height of the protrusions on a template is from about 20nm to about 40nm, or from about 40nm to about 60nm, or from about 60nm to about 80nm, or from about 80nm to about lOOnm, or from about lOOnm to about 120nm, or from about 120nm to about 140nm, or from about 140nm to about 160nm, or from about 160nm to about 180nm, or from about 180nm to about 200nm, or from about 20nm to about 200nm, or from about 40nm to about 180nm, or from about 60nm to about 140nm, or from about 80nm to about 120nm.
  • the height of the protrusions is about 20nm, or about 40nm, or about 60nm, or about 80nm, or about lOOnm, or about 120nm, or about 140nm, or about 160nm, or about 180nm, or about 200nm. In one example, the height of the protrusions is about lOOnm.
  • the protrusions on the template allows for the formation of trenches or gaps on the surface of a protein-film or a protein-metal hybrid film.
  • sub-20- nm trenches (or gaps) in the protein-metal hybrid film are able to support the localized plasmon resonance so as to confine the optical fields into the sub-20-nm trenches thereby the protein-metal hybrid film is able to function as a color filter.
  • the template comprises sub-20-nm depressions instead of protrusion
  • the protein-film or the protein-metal hybrid film manufactured using said template will comprise protrusions having width of less than 20 nm and trenches or gaps having width of more than 20 nm.
  • a fabricated protein-metal hybrid film comprising gaps having width of more than 20 nm may not be able to support the localized plasmon resonance.
  • the structures comprised on a template used in the manufacturing method described herein are of regular shape or irregular shape.
  • the irregular shapes include but are not limited to
  • the term "irregular shapes" may also refer to abstract shapes or shapes that are not considered as regular polygons.
  • Non-limiting example of the regular shape includes, but is not limited to, triangle, square, rectangle, pentagon, hexagon, octagon, any other regular polygons, and the like.
  • the structures on the template used in the method of manufacturing described herein can be faithfully replicated on the protein film.
  • the width of the structures i.e. the protrusions
  • the width of the spaces between the structures i.e. how far apart is one structure from the next
  • the structures in the template are spaced apart at from about lOnm to about 20nm, from about 20nm to about 30nm, from about 30nm to about 40nm, from about 40nm to about 50nm, from about 50nm to about 60nm, from about 60nm to about 70nm, from about 70nm to about 80nm, from about 80nm to about 90nm, from about 90nm to about lOOnm, from about lOOnm to about 120nm, from about 120nm to about 140nm, from about 140nm to about 160nm, from about 160nm to about 180nm, from about 180nm to about 200nm, from about 200nm to about 220nm, from about 220nm to about 240nm, from about 240nm to about 260nm, from about 260nm to about 280nm, from about 280nm to about 300nm, from about 300nm to about 320nm, from about 320n
  • the structures in the template are spaced apart at about lOnm, or about 20nm, or about 30nm, or about 40nm, or about 50nm, or about 60nm, or about 70nm, or about 80nm, or about 90nm, or about lOOnm, or about 120nm, or about 140nm, or about 160nm, or about 180nm, or about 200nm, or about 220nm, or about 240nm, or about 260nm, or about 280nm, or about 300nm, or about 320nm, or about 340nm, or about 360nm, or about 380nm, or about 400nm.
  • the template provided herein can be prepared according to any method known in the art, a person skilled in the art appreciate that the method to adjust the width of the spaces between the structures varies according to the method used to prepare the template.
  • the width of the protrusions on the protein- film or the protein-metal hybrid film will be determined by the width of the spaces between the structures on the template.
  • the width of the protrusion on the protein-film can also be referred as "pitch size".
  • the term "pitch size” also refers to the distance between the two nearest periodically repeated structures on the template (or protrusion on the template). Due to their sizes, the protrusions on the protein-film are considered as nanostructures. Changes in the pitch size affect the color of the protein-metal hybrid film. As shown for example on Figure 4B, increasing the pitch size causes a red shift in the reflectance optical spectrum.
  • optical reflectance spectrum refers to the reflectance values at each wavelength.
  • relative reflectance refers to the measured reflectance from one sample, with respect to the reflectance spectrum as measured from the flat region.
  • the optical reflectance spectrum of the samples can be measured using any method known in the art. In one example, the optical reflectance spectra of the samples were measured by using a CRAIC UV-VIS-NIR micro -spectrophotometer model QDI 2010 (equipped with a 36x objective lens with NA of 0.5).
  • the pitch size of a protein-metal hybrid film can be adjusted by adjusting the width of the spaces between the structures on the template used in the method of manufacturing described herein.
  • the protein solution comprises from about 0.5 wt% to about 1 wt%, or from about 1 wt% to about 1.5 wt%, or from about 1.5 wt% to about 2 wt%, or from about 2 wt% to about 2.5 wt%, or from about 2.5 wt% to about 3 wt%, or from about 3 wt% to about 3.5 wt%, or from about 3.5 wt% to about 4 wt%, or from about 4 wt% to about 4.5 wt%, or from about 4.5 wt% to about 5 wt% of protein, or from about 5 wt% to about 10 wt% of protein, or from about 10 wt% to about 15 wt% of protein, or from about 15 wt% to about 20 wt% of protein.
  • the protein solution comprises about 0.5 wt%, or about 1 wt%, or about 1.5 wt%, or about 2 wt%, or about 2.5 wt%, or about 3 wt%, or about 3.5 wt%, or about 4 wt%, or about 4.5 wt%, or about 5 wt% of protein, or about 10 wt% of protein, or about 15 wt% of protein, or about 20 wt% of protein.
  • the protein solution comprises about 0.5 wt% of protein.
  • the solvent for the peptide solution is hexafluoro-2-propanol or Hexafluoroisopropanol (HFIP).
  • the protein solution comprises a suckerin protein.
  • suckerin protein is useful for the method of manufacturing as described herein because processing of suckerin protein (e.g. dissolving suckerin protein to form protein solution) can be performed using mildly acidic solvent (such as acetic acid).
  • Non-limiting example of the use of the protein film or the protein-metal hybrid film manufactured according to the method described herein is for optical applications.
  • the template used in the method of manufacturing described herein is a template for an optical device.
  • the optical device includes, but is not limited to, a lens, a microlens array, an optical grating, a pattern generator, a beam reshaper, a color sensor, a compact spectrometer, and the like.
  • the template used in the manufacturing method described herein can be made of any material that is suitable for making a template.
  • the template used in the manufacturing method as described herein is made of a material that includes, but is not limited to, silicon, glass, metal, metal oxide, silicon dioxide, silicon nitride, indium tin oxide, ceramic, sapphire, combinations thereof, and the like.
  • the template used in the manufacturing method as described herein is made of silicon.
  • the silicon template used in the method of manufacturing described herein can be made by following any suitable protocol known in the art.
  • a non-limiting example of the protocol for formation of the silicon template is provided in Experimental Section, subtitled "Fabrication Process of Silicon Template".
  • the silicon template is formed by dry etching based on a mask defined by electron beam lithography.
  • the dry etching used for the formation of the silicon template is inductively-coupled plasma (ICP) dry etching.
  • the mask used in the dry etching process includes, but is not limited to, a mask of hydrogen silsesquioxane (HSQ), a mask of polymethyl methacrylate (PMMA) and a mask of ZEP (or ZEP resist).
  • HSQ hydrogen silsesquioxane
  • PMMA polymethyl methacrylate
  • ZEP or ZEP resist
  • the step of "allowing the proteins in the protein solution to self-assemble to form a protein film” or the step of "imprinting the protein film with a template shaped to correspond with the patterned nanostructure to form a patterned nanostructure on the protein film, wherein the template comprises structures with a width of 20 nm and below” is referred as "(b)".
  • (b) is carried out in vacuum.
  • the length of time to perform (b) is for up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, up to 6 hours, up to 7 hours, up to 8 hours, up to 9 hours, up to 10 hours, up to 12 hours, up to 14 hours, up to 16 hours, up to 18 hours, up to 20 hours, up to 22 hours, or up to 24 hours.
  • the length of time to perform (b) is more than 0 hour.
  • the length of time to perform (b) is for about 1 hour, or about 2 hours, or about 3 hours, or about 4 hours, or about 5 hours, or about 6 hours, or about 7 hours, or about 8 hours, or about 9 hours, or about 10 hours, or about 12 hours, or about 14 hours, or about 16 hours, or about 18 hours, or about 20 hours, or about 22 hours, or about 24 hours. In one example, wherein when (b) comprises the step of "allowing the proteins in the protein solution to self-assemble to form a protein film", the length of time to perform (b) is for about 6 hours.
  • the method of manufacturing described herein allows for formation of a protein film or a protein-metal hybrid film having variety of thickness.
  • the thickness of the protein film or the protein-metal hybrid film is the distance measured from surface of film that does not have protrusions to the top of the protrusions on the film.
  • the protein film is of about ⁇ to about 20 ⁇ thick, about 20 ⁇ to about 30 ⁇ thick, about 30 ⁇ to about 40 ⁇ thick, about 40 ⁇ to about 50 ⁇ thick, about 50 ⁇ to about 60 ⁇ thick, about 60 ⁇ to about 70 ⁇ thick, about 70 ⁇ to about 80 ⁇ thick, about 80 ⁇ to about 90 ⁇ thick, about 90 ⁇ to about ⁇ thick, about ⁇ to about ⁇ ⁇ thick, or about ⁇ ⁇ to about 120 ⁇ thick, or about 120 ⁇ to about 130 ⁇ thick, or about 130 ⁇ to about 140 ⁇ thick, or about 140 ⁇ to about 150 ⁇ thick, or about 150 ⁇ to about 160 ⁇ thick, or about 160 ⁇ to about 170 ⁇ thick, or about 170 ⁇ to about 180 ⁇ thick, or about 180 ⁇ to about 190 ⁇ thick, or about 190 ⁇ to about 200 ⁇ thick, or about 200 ⁇ to about 210 ⁇ thick, or about 210 ⁇ to about 220 ⁇ thick, or about 220 ⁇ to about 230 ⁇ thick, or about 230 ⁇ to about 240 ⁇ thick, or about 240 ⁇ to about 250 ⁇
  • the protein film is of ⁇ thick, or about 20 ⁇ thick, or about 30 ⁇ thick, or about 40 ⁇ thick, or about 50 ⁇ thick, or about 60 ⁇ thick, or about 70 ⁇ thick, or about 80 ⁇ m thick, or about 90 ⁇ thick, about ⁇ thick, or about ⁇ ⁇ thick, or about 120 ⁇ m thick, or about 130 ⁇ thick, or about 140 ⁇ thick, or about 150 ⁇ thick, or about 160 ⁇ thick, or about 170 ⁇ thick, or about 180 ⁇ m thick, or about 190 ⁇ thick, or about 200 ⁇ thick, or about 210 ⁇ thick, or about 220 ⁇ thick, or about 230 ⁇ m thick, or about 240 ⁇ thick, or about 250 ⁇ thick, or about 260 ⁇ thick, or about 270 ⁇ thick, or about 280 ⁇ m thick, or about 290 ⁇ thick, or about 300 ⁇ thick, or about 320 ⁇ thick, or about 340 ⁇ thick, or about 360 ⁇ m thick, or about 380 ⁇ thick, or about 400 ⁇ thick, or about 420 ⁇ thick, or about 440 ⁇ thick, or about 460
  • the method of manufacturing described herein comprises removal of a protein film from the template.
  • the template is reusable or can be re-used.
  • the multimeric design of the protein and the increase molecular weight improves the toughness of the film manufactured according to the method described herein. Therefore, when combined with a gentle lift-off process developed for demolding (i.e. removal of a protein film from the template), the structural integrity of the film is enhanced and thereby minimizing deformation in the final film.
  • removing the protein film from the template in (c) comprises lifting the protein film with mechanical tools, or allowing the protein film to attach to an adhesive material which is brought in contact with the protein film and removed after attachment of the protein film to the adhesive material, or the like.
  • the mechanical tools used for removing the protein film from the template are forceps.
  • the inventors have found that forceps are useful for removing thicker film.
  • the thickness of the protein film that can be removed using forceps is about 200 ⁇ to about 210 ⁇ thick, or about 210 ⁇ to about 220 ⁇ thick, or about 220 ⁇ to about 230 ⁇ thick, or about 230 ⁇ to about 240 ⁇ thick, or about 240 ⁇ to about 250 ⁇ thick, or about 250 ⁇ to about 260 ⁇ thick, or about 260 ⁇ to about 270 ⁇ thick, or about 270 ⁇ to about 280 ⁇ thick, or about 280 ⁇ to about 290 ⁇ thick, or about 290 ⁇ to about 300 ⁇ thick, or about 300 ⁇ to about 320 ⁇ thick, or about 320 ⁇ to about 340 ⁇ thick, or about 340 ⁇ to about 360 ⁇ thick, or about 360 ⁇ to about 380 ⁇ thick, or about 380 ⁇ to about 400 ⁇ thick, or about 400 ⁇ to about 420 ⁇ thick, or about 420 ⁇ to about
  • the thickness of the protein film that can be removed using forceps is about 200 ⁇ thick, or about 210 ⁇ thick, or about 220 ⁇ thick, or about 230 ⁇ thick, or about 240 ⁇ thick, or about 250 ⁇ thick, or about 260 ⁇ thick, or about 270 ⁇ thick, or about 280 ⁇ thick, or about 290 ⁇ thick, or about 300 ⁇ thick, or about 320 ⁇ thick, or about 340 ⁇ thick, or about 360 ⁇ thick, or about 380 ⁇ thick, or about 400 ⁇ thick, or about 420 ⁇ thick, or about 440 ⁇ thick, or about 460 ⁇ thick, or about 480 ⁇ m thick, or about 500 ⁇ thick. In one example, the thickness of the protein film that can be removed using forceps is about 300 ⁇ thick.
  • the protein film can also be removed from the template by allowing the protein film to attach to an adhesive material which is brought in contact with the protein film and removed after attachment of the protein film to the adhesive material.
  • the adhesive material used in the method of manufacturing described herein is an adhesive material that will allow for the protein-film to maintain its integrity when the adhesive material is removed. In other words, removal of the adhesive material from the protein-film will not tear, break, deform, or destroy the protein- film.
  • the adhesive material is an adhesive tape. Any type of adhesive tape that can attach to the protein film can be used in the manufacturing method described herein.
  • the adhesive tape is a dissolvable adhesive tape.
  • the manufacturing method described herein further comprising removing the adhesive tape. Any suitable method can be employed for removing the adhesive tape from the protein film.
  • removing the adhesive tape from the protein film comprises removing by mechanical force, or removing by dissolving the adhesive tape, or the like.
  • the manufacturing method described herein also comprises the step of applying a metal layer on the surface comprising patterned nanostructure of a protein-film thereby obtaining a protein-metal hybrid film.
  • the protein-metal hybrid film described herein comprises a hybrid film of protein and a metal that includes, but is not limited to, gold, silver, aluminum, titanium, chromium, platinum, copper, tin, indium, cadmium, lead, tungsten, iron, nickel, selenium, silicon, strontium, palladium, vanadium, zinc, zirconium, alloys and oxides thereof, any combination thereof, and the like.
  • the protein-metal hybrid film is a protein-gold hybrid film.
  • the step of "applying a metal layer to a surface of the protein film, wherein the surface comprises the patterned nanostructure" is referred as "(d)".
  • (d) comprises applying a gold layer to the surface of the protein film.
  • the application of the gold layer to the surface of the protein film can be performed using any application method known in the art.
  • applying a gold layer comprises evaporating gold films to the surface of the protein film.
  • the evaporation of gold films on the surface of the protein film can be performed using any suitable instrument.
  • the instrument used to evaporate gold film on the surface of protein film is Electron beam evaporator (Denton).
  • the gold films applied of the surface of the protein film can have a variety of thickness.
  • the gold films applied on the surface of the protein film are of about lnm to about 5nm thick, or about 5nm to about lOnm thick, or about lOnm to about 15nm thick, or about 15nm to about 20nm thick, or about 20nm to about 25nm thick, or about 25nm to about 30nm thick, or about 30nm to about 35nm thick, or about 35nm to about 40nm thick, or about 40nm to about 45nm thick, or about 45nm to about 50nm thick, or about 50nm to about 55nm thick, or about 55nm to about 60nm thick, or about 60nm to about 65nm thick, or about 65nm to about 70nm thick, or about 70nm to about 75nm thick, or about 75nm to about 80nm thick, or about 80nm to about 85nm thick, or about 85nm to about 90nm thick, or
  • the gold films applied on the surface of the protein film are of about lnm thick, or about 5nm thick, or about lOnm thick, or about 15nm thick, or about 20nm thick, or about 25nm thick, or about 30nm thick, or about 35nm thick, or about 40nm thick, or about 45nm thick, or about 50nm thick, or about 55nm thick, or about 60nm thick, or about 65nm thick, or about 70nm thick, or about 75nm thick, or about 80nm thick, or about 85nm thick, or about 90nm thick, or about 95nm thick, or about lOOnm thick, or about l lOnm thick, or about 120nm thick, or about 130nm thick, or about 140nm thick, or about 150nm thick, or about 160nm thick, or about 170nm thick, or about 180nm thick, or about 190nm thick, or about 200nm thick, or about 210nm thick, or about 220n
  • a protein film comprising a patterned nanostructure and a protein-metal hybrid film comprising a patterned nanostructure
  • the inventors of the present disclosure have found an alternative protein film comprising a patterned nanostructure and an alternative protein-metal hybrid film comprising a patterned nanostructure.
  • the inventors have surprisingly found that the patterned nanostructures on the protein film or on the protein-metal hybrid film are free-standing, mechanically robust, and flexible.
  • the present disclosure provides a protein film comprising a patterned nanostructure, wherein the protein is a self- assembling protein, and the patterned nanostructure has a width of 20nm and below.
  • said protein film is manufactured according to the method of manufacturing a protein film as described herein.
  • the present disclosure provides a protein-metal hybrid film comprising a patterned nanostructure, wherein the protein is a self-assembling protein, and the patterned nanostructure has a width of 20nm and below.
  • the protein- metal hybrid film is manufactured according to the method of manufacturing a protein-metal hybrid as described herein.
  • the protein film or the protein-metal hybrid film In order for the protein film or the protein-metal hybrid film to be applicable for biomedical application, it has to be biocompatible and biodegradable.
  • biocompatible refers to materials that are compatible with living tissue and/or a living system by not being toxic, injurious, or physiologically reactive, and/or not causing immunological rejection.
  • the biocompatibility of a material can be determined using any method known in the art.
  • the patterned nanostructure on the protein film as described herein or the patterned nanostructure on the protein-metal hybrid film as described herein is biocompatible.
  • the protein film as described herein or the protein- metal hybrid film as described herein is biocompatible.
  • biodegradable refers to materials that are capable of being decomposed by bacteria or other living organisms and thereby avoiding pollution.
  • the biodegradability of a material can be determined using any method known in the art.
  • the patterned nanostructure on the protein film as described herein or the patterned nanostructure on the protein-metal hybrid film as described herein is biodegradable.
  • the protein film as described herein or the protein-metal hybrid film as described herein is biodegradable.
  • the protein film described herein or the protein-metal hybrid film described herein can also comprise additional substance.
  • the present disclosure paves the way for further processing of a protein-film or a protein-metal hybrid film through incorporating with other functional elements, such as enzymes and chemical probes for biocompatible and biodegradable sensors, as well as a flexible nanoplasmonics platform.
  • other functional elements such as enzymes and chemical probes for biocompatible and biodegradable sensors, as well as a flexible nanoplasmonics platform.
  • an external heat source such as a heated plate
  • co-deposition of heat- sensitive materials such as enzymes is possible.
  • the protein film described herein or the protein-metal hybrid film described herein further comprising an additional polymer.
  • the protein film described herein or the protein-metal hybrid film described herein further comprising an active agent.
  • Any active agent that is compatible with the protein film described herein or the protein-metal hybrid film described herein can be added.
  • the active agent added to the protein film described herein or the protein-metal hybrid film described herein include, but is not limited to, therapeutic agents, cells, proteins, peptides, nucleic acid analogues, nucleotides, oligonucleotides, nucleic acids, peptide nucleic acids, aptamers, antibodies or fragments or portions thereof, hormones, hormone atagonists, growth factors or recombinant growth factors and fragments and variants thereof, cytokines, enzymes, antibiotics, antimicrobial compounds, anti-inflammation agents, antifungals, antivirals, toxins, prodrugs, chemotherapeutic agents, small molecules, dyes, amino acids, vitamins, antioxidants, combinations thereof, and the like.
  • the protein film or the protein hybrid film is useful for optical applications.
  • protein-film or hybrid protein-metal film comprising suckerins is considered as a suitable material for building optical devices.
  • Metallic nanostructures can be used to produce printed color images with resolutions beyond the diffraction limit of light.
  • the present disclosure demonstrates the patterning of suckerin protein into nanostructures with dimension of sub-20-nm, and the realization of compact optical color filters by protein-gold hybrid nanostructures. Through adjusting the pitch size and gap size of the hybrid protein-gold nanostructures, these films could be used as protein-based optical color filters, which potentially could be extended further for protein-based high-resolution color printing beyond the diffraction limit.
  • the present disclosure provides the use of the method described herein, or the protein film described herein, or the protein-metal hybrid film described herein in the manufacture of an optical device.
  • the present disclosure provides a method of manufacturing an optical device comprising attaching a protein film or a protein-metal hybrid film as defined herein to an optical device.
  • the present disclosure provides an optical device comprising a protein-metal hybrid film as defined herein.
  • the optical device comprising a protein-metal hybrid film as defined herein is an optical filter in reflection mode.
  • optical filter in reflection mode refers to an optical component, which could change or alter the reflection spectrum.
  • a protein film includes a plurality of protein films, including mixtures and combinations thereof.
  • the terms “increase” and “decrease” refer to changes of a certain measurable values such as changes in pitch size. An increase thus indicates a change on a positive scale, whereas a decrease indicates a change on a negative scale.
  • the term “change”, as used herein, also refers to the difference between a measurable value with another measurable value. However, this term is without valuation of the difference seen.
  • the term "about" in the context of concentration of a substance, thickness or width of an object (such as a protein film or a protein-metal hybrid film), length of time, or other stated values means +/- 5% of the stated value, or +/- 4% of the stated value, or +/- 3% of the stated value, or +/- 2% of the stated value, or +/- 1% of the stated value, or +/- 0.5% of the stated value.
  • range format may be disclosed in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosed ranges. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.
  • the suckerins multimer were derived from a Sepioteuthis lessoniana suckerin (suckerin- 1) that harbors both beta-sheet forming and glycine rich domains.
  • a codon- optimized gene (SEQ ID NO: 1, expressing protein as defined on SEQ ID NO: 2) for bacterial expression was ordered from IDTDNA and concatemers were generated using a one-pot reaction orthogonal end protection method.
  • Three different sets of primers (SEQ ID NOs: 3 to 8) were used to generate PCR amplicons, as corresponding to a left chain-stopping unit, middle unit and a right chain-stopping unit.
  • the left unit was digested with Bsal, the middle unit was digested with Bsal and BsmBI and the right unit was digested with BsmBI.
  • Digested amplicons were pooled at ratios of 1:2: 1, 1:4: 1 and 1:8: 1 and ligated.
  • the ligated fragments were analyzed on a 1% agarose gel and high molecular weight fragments were gel isolated, digested with Nhel and Xhol and cloned into the pET28a expression vector. Clones harbored an N-terminal HIS -tag. Insert fragments were confirmed by test digestion and sequencing.
  • Multimeric clones (SEQ ID NOs: 9, 11, 13, or 15) were transformed into T7 Express strain of E.coli cells (New England BioLabs) and streaked onto selective LB agar plates (50 ⁇ / ⁇ kanamycin) followed by an overnight incubation at 37°C. Single colonies were then inoculated into selective LB media (50 ⁇ ⁇ kanamycin) and incubated overnight under constant shaking at 37°C. Glycerol cell stocks were made by adding 20% (v/v) glycerol for storage at -80°C. Pre-cultures were made from glycerol stocks and incubated overnight.
  • Phosphate Buffered Super Broth 35g/L tryptone, 20g/L yeast extract, 2.5g/L sodium chloride, 1.12g/L potassium phosphate monobasic, 6.27g/L potassium phosphate dibasic was inoculated with the pre-culture at a ratio of 1: 100 and cultured in baffled flasks under similar conditions to an optical density (OD) of 0.4-0.6 prior to protein induction with 0.5mM IPTG.
  • OD optical density
  • Cell cultures were kept on the shaker at 37°C for another 4 hours and spun down at 4,200 rpm for 30 minutes thereafter. Cells were re-suspended twice in 50mL of 20mM tris buffer (pH 8.0) and spun down for 10 minutes at 10,000 rpm prior to storage at -20°C.
  • Multimeric clones were transformed into T7 Express strain of E.coli cells (New England BioLabs) and cultured in high density growth medium. Suckerin proteins were expressed as inclusion bodies, homogenized with high shear and purified by tangential flow filtration. The high throughput of tangential flow filtration enables purification of the proteins in large batches with excess volumes of wash buffers and minimal handling efforts. Due to a high isoelectric point of greater than pH 9, suckerin proteins are soluble in 5% acetic acid and could be processed in aqueous conditions. High fidelity replication of sub-20-nm gaps (Figure 3) onto suckerin films was thus obtained by solution casting of multi-meric suckerins under vacuum conditions, demonstrating the potential for suckerin proteins as nanopatterned substrates. When stiffer films are desired, suckerin monomer and suckerin trimer were used. Pentamer was largely used for its high yield and robustness. [0068] Fabrication Process of Silicon Template.
  • the sample was immediately rinsed by acetone and isopropanol alcohol (IPA), followed by a continuous flow of nitrogen air gun for 1 minute to dry the sample.
  • IPA isopropanol alcohol
  • the next process is silicon etching by using an inductively-coupled-plasma (ICP, Unaxis shuttle lock system SLR-7701-BR), where the detailed etching conditions were DC power of 150 watts, coil power of 300 watts, Cl 2 /HBr with the flow rates of 18 seem and 22 seem, process pressure of 10 mtorr, temperature at 6 °C, and an etching time of 46 seconds.
  • ICP inductively-coupled-plasma
  • Suckerins are composed of a family of modular proteins with a distinct architecture that facilitates the formation of beta- sheet-reinforced polymer networks.
  • Thirty eight (38) different suckerins from squid and cuttlefish that are part of a multigene family were recently identified.
  • a distinct feature of suckerin is that they are composed of a series of repetitive beta-sheet forming modules (Ml) and amorphous Gly-rich modules (M2) with precise placement of proline residues, which constrain beta-sheet nanocrystal size.
  • Ml beta-sheet forming modules
  • M2 amorphous Gly-rich modules
  • Each of the 38 different suckerin proteins has distinct molecular weights and molecular architectures.
  • a simple casting procedure to produce suckerin films from nano-patterned silicon substrates with sub-20-nm features was developed.
  • the silicon templates were patterned by the dry etching process of using inductively-coupled plasma (ICP), and the etching mask was HSQ pattern as defined by electron beam lithography (EBL).
  • EBL electron beam lithography
  • the silicon substrate was treated by trichloro (lH,lH,2H,2H-perfluorooctyl) silane. Lyophilized pentamer was dissolved to a concentration of 5 mg/mL in 0.22 ⁇ filtered 5% acetic acid and vacuum casted for 6 hours onto the silicon template as shown in Figure 2B.
  • FIG. 3 presents the scanning electron micrograph (SEM) characterization results of the patterned protein nanostructures with various shapes. These SEM images of protein nanostructures were taken after evaporating 2-nm-thick Chromium (i.e. Cr) so as to avoid the charging effect during SEM imaging. It shows that the typical gap size of the patterned protein nanostructures is less than 20 nm. These characterization results demonstrate that suckerin protein could be used as a simple high resolution patterning resist with the advantage of bio-compatibility, as compared to the existing approach by using gold and optical adhesive (OA) glue.
  • SEM scanning electron micrograph
  • FIG. 4A shows the corresponding observable color of the features with varying pitch and varying dosage
  • Figure 4B presents the measured optical reflectance spectrum.
  • dose or "dose time” as used herein refers to the time that electron beam lithography exposes the HSQ resist. The length of the "dose time” can be controlled using the software interface of the electron beam lithography instrument.
  • SEM of Feature 9 ( Figure 4C) serves as evidence of the sub-20-nm gap size that is present in the features even after the gold evaporation process.

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Abstract

La présente invention concerne un procédé de fabrication d'un film de protéine comprenant une nanostructure à motifs, la protéine étant une protéine à auto-assemblage, le procédé comprenant : (a) appliquer une solution de protéine à un modèle formé correspondant à la nanostructure à motifs, le modèle comprenant des structures ayant une largeur inférieure à 20 nm ; (b) permettre aux protéines de la solution protéique de s'auto-assembler afin de former un film protéique ; et (c) retirer le film protéique du modèle. La présente invention concerne également des procédés de fabrication de films hybrides de protéine-métal et des films de protéine résultants et des films hybrides de protéine-métal de ceux-ci. Dans un mode de réalisation préféré, la protéine est une protéine suckérine (protéine de couronne de ventouse), et les films hybrides protéine-métal sont utilisés dans des dispositifs optiques.
PCT/SG2017/050516 2016-10-13 2017-10-13 Formation directe de motifs en dessous de 20 nm de protéine recombinante par fabrication de transfert de motif Ceased WO2018070942A1 (fr)

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WO2009061823A1 (fr) * 2007-11-05 2009-05-14 Trustees Of Tufts College Fabrication de structures photoniques de fibroïne de soie par impression par nanocontact
WO2014062134A1 (fr) * 2012-10-17 2014-04-24 Nanyang Technological University Composés et procédés pour la production de suckérine et utilisations de ceux-ci
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WO2009061823A1 (fr) * 2007-11-05 2009-05-14 Trustees Of Tufts College Fabrication de structures photoniques de fibroïne de soie par impression par nanocontact
WO2014062134A1 (fr) * 2012-10-17 2014-04-24 Nanyang Technological University Composés et procédés pour la production de suckérine et utilisations de ceux-ci
WO2014160131A1 (fr) * 2013-03-14 2014-10-02 The Penn State Research Foundation Compositions et procédés associés à des protéines aptes à une transition réversible en un produit de fusion

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RIEU C. ET AL.: "The role of water on the structure and mechanical properties of a thermoplastic natural block co-polymer from squid sucker ring teeth", BIOINSPIR BIOMIM., vol. 11, no. 5, 2 September 2016 (2016-09-02), pages 1 - 10, XP020309237, [retrieved on 20171214] *

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