WO2011113592A1 - Procédé de fabrication de fibres contenant un polymère - Google Patents

Procédé de fabrication de fibres contenant un polymère Download PDF

Info

Publication number
WO2011113592A1
WO2011113592A1 PCT/EP2011/001307 EP2011001307W WO2011113592A1 WO 2011113592 A1 WO2011113592 A1 WO 2011113592A1 EP 2011001307 W EP2011001307 W EP 2011001307W WO 2011113592 A1 WO2011113592 A1 WO 2011113592A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
module
fibre
repetitive
amino acid
Prior art date
Application number
PCT/EP2011/001307
Other languages
English (en)
Inventor
Andreas Bausch
Sebastian Rammensee
Original Assignee
Amsilk Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Amsilk Gmbh filed Critical Amsilk Gmbh
Priority to AU2011229482A priority Critical patent/AU2011229482B2/en
Priority to CA2789647A priority patent/CA2789647C/fr
Priority to RU2012144053/05A priority patent/RU2545331C2/ru
Priority to US13/634,484 priority patent/US20130172478A1/en
Priority to EP11709018.3A priority patent/EP2547810B1/fr
Publication of WO2011113592A1 publication Critical patent/WO2011113592A1/fr

Links

Classifications

    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F1/00—General methods for the manufacture of artificial filaments or the like
    • D01F1/02—Addition of substances to the spinning solution or to the melt
    • 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
    • C07K14/43563—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
    • D01D5/00—Formation of filaments, threads, or the like
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F4/00—Monocomponent artificial filaments or the like of proteins; Manufacture thereof
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F6/00—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof
    • D01F6/58—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from homopolycondensation products
    • D01F6/68—Monocomponent artificial filaments or the like of synthetic polymers; Manufacture thereof from homopolycondensation products from polyaminoacids or polypeptides
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T442/00—Fabric [woven, knitted, or nonwoven textile or cloth, etc.]
    • Y10T442/30—Woven fabric [i.e., woven strand or strip material]
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T442/00—Fabric [woven, knitted, or nonwoven textile or cloth, etc.]
    • Y10T442/60—Nonwoven fabric [i.e., nonwoven strand or fiber material]

Definitions

  • the present invention relates to a method of spinning a polymer fibre. It further relates to a polymer fibre obtainable by said method and to uses thereof. The invention also relates to products comprising said polymer fibre.
  • Natural spider silk can assume different forms, depending on the gland it is produced in (Gosline et al., J. Exp. Biol. (202):3295, 1999). Its properties are remarkable: Its tensile strength can be superior to that of steel and equals that of aramid filaments, e.g. Kevlar. Spider silk can also be very ductile, being stretchable to up to about 300% of its length without tearing. Above all, it is lightweight.
  • Spinning of fibres is a process in which deforming stresses in the direction of the fibre axis compete with surface tension of the spinning dope. For example, it is not possible to pull out a stable fluid filament of water due to the high surface tension of water.
  • the Ohnesorge number Oh describes the ratio of viscous to surface tension forces.
  • Recombinant spider silk protein can be produced by bacteria and its assembly has been studied in detail (Scheibel, Microb. Cell. Fact., (1 ): 14, 2004; Huemmerich et al., Curr. Biol., (22):2070-4, 2004; Rammensee et al, PNAS, (105): 6590-6595, 2008).
  • Artificial spinning dopes with low protein concentrations (10-20 mg/ml eADF3 and eADF4) do hot show Tr »1.
  • the surface tension is still much larger than the viscosity effects, so also Oh « 1, therefore, fibre formation from these low protein concentration solutions has not been possible.
  • the inventors of the present invention surprisingly found that only very low polymer concentrations are required for spinning a fibre with the method of the present invention.
  • the use of very low polymer concentrations has the advantage, that the shear viscosity of the solution is not increased, which, in turn, improves the processability of the fluid.
  • fibre formation from very low concentrated polymers e.g. silk polypeptides such as spider silk polypeptides, under gentle conditions was facilitated by the inclusion of elongational viscosity enhancers.
  • spider silk dopes in nature have been described as containing very high percentages of spider silk protein, the present method surprisingly requires only very low concentrations of silk polypeptides beginning at concentrations as low as 0.15 mg/ml. .
  • the present method surprisingly requires only very low concentrations of the above-mentioned elongational viscosity enhancers.
  • the low concentrations of said enhancers are sufficient to increase on the one hand side the elongational viscosity of the solution, but on the other hand side do not substantially increase the shear viscosity of the solution so that the generation of fibres is improved.
  • the use of very low concentrations of polymers and/or very low concentrations of elongational viscosity enhancers in the method of the present invention facilitates the spinning of fibres from a spinning solution. Furthermore, the method has a low technical hurdle with fibres being drawn directly from the silk polypeptide solution at ambient or near ambient conditions.
  • the present invention relates to a method for fibre spinning from a spinning solution comprising the steps of:
  • a spinning solution comprising (i) a polymer that can be brought in to an aqueous solution in a concentration of at least 0.15 mg/ml, (ii) a compound that increases elongational viscosity of the spinning solution and (iii) a solvent; and
  • the present invention relates to a spinning solution for carrying out the method of the present invention, comprising (i) a polymer that can be brought in to an aqueous solution in a concentration of at least 0.15 mg/ml, (ii) a compound that increases elongational viscosity of the spinning solution and (iii) a solvent.
  • the present invention relates to a fibre obtainable by the method of the first aspect and to its use.
  • the present invention provides products comprising the fibre of the second aspect.
  • the terms used herein are defined as described in "A multilingual glossary of biotechnological terms: (IUPAC Recommendations)", Leuenberger, H.G.W, Nagel, B. and olbl, H. eds. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland).
  • Residues in two or more polypeptides are said to "correspond" to each other if the residues occupy an analogous position in the polypeptide structures. It is well known in the art that analogous positions in two or more polypeptides can be determined by aligning the polypeptide sequences based on amino acid sequence or structural similarities. Such alignment tools are well known to the person skilled in the art and can be, for example, obtained on the World Wide Web, e.g., ClustalW (www.ebi.ac.uk/clustalw) or Align (http ://www. ebi . ac.uk/emboss/ ali gn/index .html) using standard settings, preferably for Align EMBOSS: needle, Matrix: Blosum62, Gap Open 10.0, Gap Extend 0.5.
  • ClustalW www.ebi.ac.uk/clustalw
  • Align http ://www. ebi . ac.uk/emboss
  • the present invention relates to a method of spinning a fibre from a spinning solution comprising, essentially consisting of, or consisting of the steps of:
  • a spinning solution comprising (i) a polymer that can be brought in to an aqueous solution in a concentration of at least 0.15 mg/ml, (ii) a compound that increases elongational viscosity of the spinning solution and (iii) a solvent; and
  • polymer refers to a molecule composed of repeating structural units typically connected by covalent chemical bonds.
  • the polymer is a biopolymer such as a polypeptide or a synthetic polymer.
  • biopolymer refers to a molecule containing monomelic units that are covalently bonded to form larger structures. Biopolymers can be classified in three main classes based on the differing monomeric units used and the structure of the biopolymers formed.
  • the first class of biopolymers are polynucleotides which are polymers composed of nucleotide monomers, e.g. polymers composed of 10 or more nucleotide monomers.
  • the second class of biopolymers are polypeptides which are polymers of amino acids.
  • the third class of biopolymers are polysaccharides which are often linear bonded polymeric carbohydrate structures. It is preferred that the biopolymer is a polypeptide.
  • the biopolymer is biodegradable and more preferably also compostable.
  • Biodegradable biopolymers are broken down into C0 2 and water by microorganisms.
  • Biodegradable biopolymers which are also compostable can be put into an industrial composting process and will break down within months.
  • biopolymer is biocompatible. This means that the polymer is non-toxic, non-mutagenic and elicits no or only a minimal inflammatory reaction.
  • fibres spun from biopolymers with the method of the present invention can have the above mentioned advantageously characteristics, namely can be biodegradable, compostable and/or biocompatible or can be mostly biodegradable, compostable and/or biocompatible.
  • the amount of elongational viscosity enhancers is preferably very low in the spinning solution of the method of the present invention.
  • the repeating structural units are amino acids connected by covalent amide bonds (peptide bonds).
  • polypeptide and “protein” are used interchangeably herein and mean any peptide-linked chain of amino acids, regardless of length or post-translational modification.
  • the polypeptide is a silk polypeptide comprising at least two identical repetitive units, bovine serum albumin (BSA), zein or casein.
  • BSA bovine serum albumin
  • the polypeptide may also be collagen, fibronectin or keratin.
  • Bovine serum albumin also known as “Fraction V”
  • Fraction V is a serum albumin protein.
  • Zein is a prolamine protein found in maize.
  • Casein from Latin caseus “cheese”
  • aSl phosphoprotein
  • aS2 phosphoprotein
  • ⁇ phosphoprotein
  • the term "silk polypeptide” refers to a silk polypeptide or protein (it is noted that, unless otherwise indicated, these two terms, as used herein, are interchangeable) that is expressed in a recombinant (e.g. microbial, yeast, plant, insect or mammalian) expression system, i.e. separated from its natural milieu in the spider gland (recombinant silk polypeptide or protein).
  • a recombinant silk polypeptide or protein i.e. separated from its natural milieu in the spider gland (recombinant silk polypeptide or protein).
  • the "silk polypeptide” is isolated or purified.
  • a “purified silk polypeptide” or an “isolated silk polypeptide” is free or substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the protein is isolated or derived.
  • substantially free of cellular material includes preparations of a silk polypeptide in which the silk polypeptide is separated from cellular components of the cells from which it is recombinantly produced.
  • a silk polypeptide that is substantially free of cellular material includes preparations of silk polypeptides having less than about 30%, 20%, 10%, 5% or 1 % (by dry weight) of contaminating protein.
  • the silk polypeptide is expressed in cell culture, it is also free or substantially free of culture medium, i.e., the culture medium represents less than about 20%, 10%, 5% or 1 % of the volume of the polypeptide preparation.
  • a "silk polypeptide” as used in the context of the present invention further refers to a polypeptide with an amino acid sequence which comprises or consists of at least 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, preferably at least 95% and most preferably 100% of multiple copies of one identical repetitive unit (e.g. A 2 , Q 6 , or C 1 , wherein the items 2, 6, or 16 represent the number of repetitive units) or multiple copies of two or more different repetitive units (e.g. (AQ) 24 , or (AQ) 12 , Ci 6 ).
  • one identical repetitive unit e.g. A 2 , Q 6 , or C 1 , wherein the items 2, 6, or 16 represent the number of repetitive units
  • two or more different repetitive units e.g. (AQ) 24 , or (AQ) 12 , Ci 6 .
  • silk polypeptide also refers to a silk polypeptide that comprises or consists of at least two identical repetitive units which comprise or consists of identical copies of amino acid sequences of naturally- occurring silk polypeptides or of variations of amino acid sequences of naturally- occurring silk polypeptides or of combinations of both.
  • a “repetitive unit” refers to a region which corresponds in amino acid sequence to a region that comprises or consists of at least one peptide motif (e.g. AAAAAA (SEQ ID NO: 13) or GPGQQ (SEQ ID NO: 4)) that repetitively occurs within a naturally occurring silk polypeptide (e.g. MaSpI, ADF- 3, ADF-4, or Flag) (i.e. identical amino acid sequence) or to an amino acid sequence substantially similar thereto (i.e. variational amino acid sequence).
  • a naturally occurring silk polypeptide e.g. MaSpI, ADF- 3, ADF-4, or Flag
  • substantially similar means a degree of amino acid identity of at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or even 99.9%, preferably over the whole length of the respective reference naturally occurring amino acid sequence.
  • a “repetitive unit” having an amino acid sequence which is "substantially similar” to a corresponding amino acid sequence within a naturally occurring silk polypeptide is also similar with respect to its functional properties, e.g. a silk polypeptide comprising the "substantially similar repetitive unit” still has the ability to form a fibre.
  • a silk polypeptide comprising the "substantially similar repetitive unit” still has the ability to form a fibre.
  • the skilled person can visually assess whether a fibre is still formed.
  • the smooth and homogenous appearance of said fibre can be controlled using electronic-microscopy.
  • a “repetitive unit" having an amino acid sequence which is “identical” to the amino acid sequence of a naturally occurring silk polypeptide for example, can be a portion of a silk polypeptide corresponding to one or more peptid motifs of MaSp I (SEQ ID NO: 43) MaSp II (SEQ ID NO: 44), ADF-3 (SEQ ID NO: 1) and/or ADF-4 (SEQ ID NO: 2).
  • a “repetitive unit" having an amino acid sequence which is "substantially similar" to the amino acid sequence of a naturally occurring silk polypeptide for example, can be a portion of a silk polypeptide corresponding to one or more peptide motifs of MaSpI (SEQ ID NO: 43) MaSpII (SEQ ID NO: 44), ADF-3 (SEQ ID NO: 1) and or ADF-4 (SEQ ID NO: 2), but having one or more amino acid substitutions at specific amino acid positions.
  • the “repetitive unit” does not include the non-repetitive hydrophilic amino acid domains generally thought to be present at the carboxy and amino terminals of naturally occurring silk polypeptides.
  • a “repetitive unit” according to the present invention further refers to an amino acid sequence with a length of 3 to 200 amino acids, or 5 to 150 amino acids, preferably with a length of 10 to 100 amino acids, or 15 to 80 amino acids and more preferably with a length of 18 to 60, or 20 to 40 amino acids.
  • the repetitive unit according to the present invention can have a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180,
  • the silk polypeptide as used in the method of the present invention preferably consists of between 6 to 1,500 amino acids, between 50 to 1,500 amino acids, or between 200 to 1,300 amino acids and most preferably between 250 to 1,200 amino acids, or between 500 to 1,000 amino acids.
  • the silk polypeptide used in the method of the present invention comprises or consists of between 2 to 80 repetitive units, between 3 to 80 repetitive units, or between 4 to 60 repetitive units, more preferably between 8 to 48 repetitive units, or between 10 to 40 repetitive units and most preferably between 16 to 32 repetitive units.
  • the silk polypeptide used in the method of the present invention can comprise or consists of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79 or 80 repetitive units.
  • the silk polypeptide comprises 4, 8, 12, 16, 24, 32 or 48 repetitive units.
  • the silk polypeptide used in the method of the present invention may comprise multiple copies of one identical repetitive unit (e.g. A 2 or Ci 6 , wherein the items 2 or 6 represent the number of repetitive units) or multiple copies of two or more different repetitive units (e.g. (AQ) 24 or (QAQ) 8 ).
  • the silk polypeptide used in the method of the present invention can comprise or consist of an amino acid sequence of any silk polypeptide known to one skilled in the art. It is preferred that the silk polypeptide used in the method of the present invention comprises or consists of an amino acid sequence of an insect silk polypeptide, preferably of an arthropod polypeptide, or a spider silk polypeptide. The silk polypeptide used in the method of the present invention can also comprise or consist of an amino acid sequence of a mussel silk polypeptide.
  • the spider silk polypeptide comprises or consists of an amino acid sequence of a major ampullate gland polypeptide (MaSp), such as a dragline spider silk polypeptide, a minor ampullate gland polypeptide (MiSp), a flagelliform polypeptide, an aggregate spider silk polypeptide, an aciniform spider silk polypeptide or a pyriform spider silk polypeptide.
  • the spider silk polypeptide comprises or consists of an amino acid sequence of a dragline spider silk polypeptide or a flagelliform spider silk polypeptide. It is generally preferred to select the amino acid sequence of the dragline polypeptide or flagelliform polypeptide of a dragline polypeptide or flagelliform polypeptide of orb-web spiders of Araneidae or Araneoids.
  • the insect silk polypeptide comprises or consists of an amino acid sequence of a silk polypeptide of Lepidoptera. More preferably, the insect silk polypeptide comprises or consists of an amino acid sequence of a silk polypeptide of Bombycidae, most preferably of Bombyx mori.
  • the above mentioned silk polypeptides are recombinantly produced, i.e. are recombinant silk polypeptides.
  • the silk polypeptides used in the method of the present invention are recombinant spider silk polypeptides such as dragline spider silk polypeptides or flagelliform spider silk polypeptides, recombinant insect silk polypeptides, or recombinant mussel silk polypeptides.
  • the repetitive unit of the silk polypeptide used in the method of the present invention can comprise or consist of an amino acid sequence of any region that comprises or consists of at least one peptide motif that repetitively occurs within a naturally occurring silk polypeptide known to one skilled in the art.
  • the repetitive unit of the silk polypeptide used in the method of the present invention comprises or consists of an amino acid sequence of a region that comprises or consists of at least one peptide motif that repetitively occurs within an insect silk polypeptide, more preferably within an arthropod silk polypeptide or a spider silk polypeptide.
  • the repetitive unit of the silk polypeptide used in the method of the present invention can also comprise or consist of an amino acid sequence of a region that comprises or consists of at least one peptide motif that repetitively occurs within a mussel silk polypeptide.
  • the spider silk repetitive unit comprises or consists of an amino acid sequence of a region that comprises or consists of at least one peptide motif that repetitively occurs within a naturally occurring major ampullate gland polypeptide (MaSp), such as a dragline spider silk polypeptide, a minor ampullate gland polypeptide (MiSp), a flagelliform polypeptide, an aggregate spider silk polypeptide, an aciniform spider silk polypeptide or a pyriform spider silk polypeptide.
  • the repetitive unit comprises or consists of an amino acid sequence of a region that comprises or consists of at least one peptide motif that repetitively occurs within a naturally occurring dragline spider silk polypeptide or a flagelliform spider silk polypeptide.
  • the insect silk repetitive unit comprises or consists of an amino acid sequence of a region that comprises or consists of at least one peptide motif that repetitively occurs within a naturally occurring silk polypeptide of Lepidoptera. More preferably, the insect silk repetitive unit comprises or consists of an amino acid sequence of a region that comprises or consists of at least one peptide motif that repetitively occurs within a naturally occurring insect silk polypeptide of Bombycidae, most preferably of Bombyx mori.
  • Consensus sequence refers to an amino acid sequence which contains amino acids which frequently occur in a certain position (e.g. "G") and wherein, other amino acids which are not further determined are replaced by the place holder "X".
  • the silk polypeptide used in the method of the present invention comprises, essentially consists of, or consists of at least two identical repetitive units each comprising at least one, preferably one, consensus sequence selected from the group consisting of:
  • GPGXX (SEQ ID NO: 3), wherein X is any amino acid, preferably in each case independently selected from A, S, G, Y, P, N and Q;
  • GGX GGX
  • X is any amino acid, preferably in each case independently selected from Y, P, R, S, A, T, N and Q, more preferably in each case independently selected from Y, P and Q; and iii) A x , wherein x is an integer from 5 to 10.
  • the silk polypeptide used in the method of the present invention comprises or consists of at least two identical repetitive units each comprising at least one, preferably one amino acid sequence selected from the group consisting of: GGRPSDTYG (SEQ ID NO: 18) and GGRPSSSYG (SEQ ID NO: 19).
  • the iterated (peptide) motifs GPGXX (SEQ ID NO: 3) and GGX i.e. glycine rich motifs, provide flexibility to the silk polypeptide and thus, to the thread formed from the silk protein containing said motifs.
  • the iterated GPGXX (SEQ ID NO: 3) motif forms ⁇ -turn spiral structures, which imparts elasticity to the silk polypeptide.
  • Major ampullate and flagelliform silks both have a GPGXX (SEQ ID NO: 3) motif.
  • the iterated GGX motif is associated with a helical structure having three amino acids per turn and is found in most spider silks. The GGX motif may provide additional elastic properties to the silk.
  • the iterated polyalanine A x (peptide) motif forms a crystalline ⁇ -sheet structure that provides strength to the silk polypeptide.
  • the GGRPSDTYG (SEQ ID NO: 18) and GGRPSSSYG (SEQ ID NO: 19) (peptide) motifs have been selected from Resilin (WO 08/155304).
  • Resilin is an elastomeric protein found in most arthropods (arthropoda). It is located in specialised regions of the cuticle, providing low stiffness and high strength (Elvin et al., Nature (473): 999-1002, 2005).
  • the silk polypeptide comprises or consists- of repetitive units each comprising at least one (e.g. 1, 2, 3, 4, 5, 6, 7, 8, or 9), preferably one, amino acid sequence selected from the group consisting of GPGAS (SEQ ID NO: 5), GPGSG (SEQ ID NO: 6), GPGGY (SEQ ID NO: 7), GPGGP (SEQ ID NO: 8), GPGGA (SEQ ID NO: 9), GPGQQ (SEQ ID NO: 4), GPGGG (SEQ ID NO: 10), GPGQG (SEQ ID NO: 40), and GPGGS (SEQ ID NO: 1 1).
  • GPGAS SEQ ID NO: 5
  • GPGSG SEQ ID NO: 6
  • GPGGY SEQ ID NO: 7
  • GPGGP SEQ ID NO: 8
  • GPGGA SEQ ID NO: 9
  • GPGQQ SEQ ID NO: 4
  • GPGGG SEQ ID NO: 10
  • GPGQG SEQ ID NO: 40
  • GPGGS SEQ ID NO:
  • the silk polypeptide comprises or consists of repetitive units each comprising at least one (e.g. 1, 2, 3, 4, 5, 8, 7, or 8), preferably one, amino acid sequence selected from the group consisting of GGY, GGP, GGA, GGR, GGS, GGT, GGN, and GGQ.
  • the silk polypeptide comprises or consists of repetitive units each comprising at least one (e.g.
  • AAAAA amino acid sequence selected from the group consisting of AAAAA (SEQ ID NO: 12), AAAAAA (SEQ ID NO: 13), AAAAAAA (SEQ ID NO: 14), AAAAAAAA (SEQ ID NO: 15), AAAAAAAAA (SEQ ID NO: 16), and AAAAAAAAAA (SEQ ID NO: 17).
  • the silk polypeptide comprises or consists of repetitive units each comprising at least one (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25), preferably one, amino acid sequence selected from the group consisting of GPGAS (SEQ ID NO: 5), GPGSG (SEQ ID NO: 6), GPGGY (SEQ ID NO: 7), GPGGP (SEQ ID NO: 8), GPGGA (SEQ ID NO: 9), GPGQQ (SEQ ID NO: 4), GPGGG (SEQ ID NO: 10), GPGQG (SEQ ID NO: 40), GPGGS (SEQ ID NO: 11), GGY, GGP, GGA, GGR, GGS, GGT, GGN, GGQ, AAAAA (SEQ ID NO: 12), AAAAAA (SEQ ID NO: 13), AAAAAAA (SEQ ID NO: 14), AAAAAAAA (SEQ ID NO: 15), AAAAAAAAA (SEQ ID NO: 16
  • the silk polypeptide used in the method of the present invention comprises, essentially consist of, or consists of repetitive units, which comprise or consist of
  • GPGAS SEQ ID NO: 5
  • AAAAAA SEQ ID NO: 13
  • GGY GGGY
  • GPGSG SEQ ID NO: 6
  • AAAAAAAA SEQ ID NO: 15
  • GPGGY SEQ ID NO: 7
  • GPGGY SEQ ID NO: 7
  • GPGGP SEQ ID NO: 8
  • GPGGA SEQ ID NO: 9
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GGP as amino acid sequence, preferably in this order
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GGP as amino acid sequence, preferably in this order
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GGP as amino acid sequence, preferably in this order
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GGP as amino acid sequence, preferably in this order
  • GGP GPGGA (SEQ ID NO: 9)
  • GGP GGP as amino acid sequence, preferably in this order
  • AAAAAAAA SEQ ID NO: 15
  • GPGGG SEQ ID NO: 10
  • GGR GGN
  • GGR GGN
  • the silk polypeptide used in the method of the present invention comprises, essentially consists of, or consists of between 2 to 80 repetitive units, between 3 to 80 repetitive units, or between 4 to 60 repetitive units, more preferably between 8 to 48 repetitive units, or between 10 to 40 repetitive units and most preferably between 16 to 32 repetitive units, i.e.
  • GPGXX (SEQ ID NO: 3), wherein X is any amino acid, preferably in each case independently selected from A, S, G, Y, P, N and Q;
  • GGX GGX
  • X is any amino acid, preferably in each case independently selected from Y, P, R, S, A, T, N and Q, more preferably in each case independently selected from Y, P and Q; and iii) A x , wherein x is an integer from 5 to 10.
  • At least two of the repetitive units comprised in the silk polypeptide used in the method of the present invention are identical repetitive units.
  • the silk polypeptide used in the method of the present invention comprises or consists of between 2 to 80 repetitive units, between 3 to 80 repetitive units, or between 4 to 60 repetitive units, more preferably between 8 to 48 repetitive units, or between 10 to 40 repetitive units and most preferably between 16 to 32 repetitive units, each comprising at least one, preferably one amino acid sequence selected from the group consisting of: GGRPSDTYG (SEQ ID NO: 18) and GGRPSSSYG (SEQ ID NO: 19).
  • the silk polypeptide used in the method of the present invention preferably comprises, essentially consists of, or consists of between 2 to 80 repetitive units, between 3 to 80 repetitive units, or between 4 to 60 repetitive units, more preferably between 8 to 48 repetitive units, or between 10 to 40 repetitive units and most preferably between 16 to 32 repetitive units, each comprising at least one (e.g.
  • GPGAS GPGAS
  • GPGSG SEQ ID NO: 6
  • GPGGP SEQ ID NO: 8
  • GPGGA SEQ ID NO: 9
  • GPGQQ SEQ ID NO: 4
  • GPGQG SEQ ID NO: 40
  • GPGGG SEQ ID NO: 10
  • GPGGS SEQ ID NO: 11
  • GGY GGP
  • GGA GGR
  • GGS GGT
  • GGN GGN
  • GGQ AAAAA
  • AAAAA SEQ ID NO: 12
  • AAAAAA SEQ ID NO: 13
  • AAAAAAA SEQ ID NO: 14
  • AAAAAAAA SEQ ID NO: 15
  • AAAAAAAAA SEQ ID NO: 16
  • AAAAAAAAAAAA SEQ ID NO: 17
  • GGRPSDTYG SEQ ID NO: 18
  • the silk polypeptide used in the method of the present invention comprises, essentially consists of, or consists of
  • repetitive units which comprise or consist of GPGAS (SEQ ID NO: 5), AAAAAA (SEQ ID NO: 13), GGY, and GPGSG (SEQ ID NO: 6) as amino acid sequence, preferably in this order,
  • repetitive units which comprise or consist of GPGQQ (SEQ ID NO: 4), GPGQQ (SEQ ID NO: 4), GPGQQ (SEQ ID NO: 4) and GPGQQ (SEQ ID NO: 4) as amino acid sequence, preferably in this order,
  • repetitive units which comprise or consist of GPGGA (SEQ ID NO: 9), GGP, GPGGA (SEQ ID NO: 9), GGP, GPGGA (SEQ ID NO: 9), and GGP as amino acid sequence, preferably in this order
  • repetitive units which comprise or consist of AAAAAAAA (SEQ ID NO: 15), GPGQG (SEQ ID NO: 40), and GGR as amino acid sequence, preferably in this order
  • GPGGG SEQ ID NO: 10
  • GGR GGN
  • GGR amino acid sequence
  • repetitive units which comprise or consist of GGA, GGA, GGA, GGS, GGA, and GGS as amino acid sequence, preferably in this order, and/or
  • repetitive units which comprise or consist of GPGGA (SEQ ID NO: 9), GPGGY (SEQ ID NO: 7), GPGGS (SEQ ID NO: 11), GPGGY (SEQ ID NO:
  • the silk polypeptide used in the method of the present invention comprises, essentially consist of, or consists of
  • GGXn ii) (GGX)n as a repetitive unit, wherein X is any amino acid, preferably in each case independently selected from Y, P, R, S, A, T, N and Q, more preferably in each case independently selected from Y, P and Q, and n is 2, 3, 4, 5, 6, 7, or 8; and/or
  • x is an integer from 5 to 10 and n is 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • At least two of the repetitive units comprised in the silk polypeptides used in the method of the present invention are identical repetitive units.
  • the repetitive units are independently selected from module A (SEQ ID NO: 20), module C (SEQ ID NO: 21), module Q (SEQ ID NO: 22), module K (SEQ ID NO: 23), module sp (SEQ ID NO: 24), module S (SEQ ID NO: 25), module R (SEQ ID NO: 26), module X (SEQ ID NO: 27), or module Y (SEQ ID NO: 28), or variants thereof (i.e. module A variants, module C variants, module Q variants, module K variants, module sp variants, module S variants, module R variants, module X variants or module Y variants).
  • the modules A (SEQ ID NO: 20) and Q (SEQ ID NO: 22) are based on the amino acid sequence of ADF-3 of the spider Araneus diadematus.
  • Module C (SEQ ID NO: 21) is based on the amino acid sequence of ADF-4 of the spider Araneus diadematus.
  • the modules K (SEQ ID NO: 23), sp (SEQ ID NO: 24), X (SEQ ID NO: 27) and Y (SEQ ID NO: 28) are based on the amino acid sequence of the flagelliform protein FLAG of the spider Nephila clavipes (WO 2006/008163).
  • the modules S (SEQ ID NO: 25) and R (SEQ ID NO: 26) are based on Resilin (Arthropoda) (WO 2008/155304).
  • the repetitive units of the silk polypeptide consist of module A: GPYGPGASAAAAAAAAGGYGPGSGQQ (SEQ ID NO: 20), module C: GSSAAAAAAAAS ' GPGGYGPENQGPSGPGGYGP GGP (SEQ ID NO: 21), module Q: GPGQQGPGQQGPGQQQ (SEQ ID NO: 22), module K: GPGGAGGPYGPGGAGGPYGPGGAGGPY (SEQ ID NO: 23), module sp: GGTTIIEDLDITIDGADGPITISEELTI (SEQ ID NO: 24), module S: PGSSAAAAAAAAAASGPGQGQGQGQGGRPSDTYG (SEQ ID NO: 25), module R: SAAAAAAAAGPGGGNGGRPSDTYGAPGGGNGGRPSSSYG (SEQ ID NO: 26), module X: GGAGGAGGAGGSGGAGGS (SEQ ID NO: 27), or
  • the silk polypeptide used in the method of the present invention comprises, essentially consists of, or consists of between 2 o 80 repetitive units, between 3 to 80 repetitive units, or between 4 to 60 repetitive units, more preferably between 8 to 48 repetitive units, or between 10 to 40 repetitive units and most preferably between 16 to 32 repetitive units, i.e.
  • module A variants module C variants, module Q variants, module K variants, module sp variants, module S variants, module R variants, module X variants or module Y variants). It should be noted that at least two of the repetitive units comprised in the silk polypeptide used in the method of the present invention are identical repetitive units (modules).
  • the silk polypeptide used in the method of the present invention comprises, essentially consists of, or consists of (i) repetitive unit(s) consisting of module A and/or repetitive unit(s) consisting of module A variants, (ii) repetitive unit(s) consisting of module C and/or repetitive unit(s) consisting of module C variants, (iii) repetitive unit(s) consisting of module Q and/or repetitive unit(s) consisting of module Q variants, (iv) (a) repetitive unit(s) consisting of module A and repetitive unit(s) consisting of module Q, (b) repetitive unit(s) consisting of module A and repetitive unit(s) consisting of module Q variants, (c) repetitive unit(s) consisting of module A variants and repetitive unit(s) consisting of module Q, (d) repetitive unit(s) consisting of module A variants and repetitive unit(s) consisting of module Q variants, (v) (a) repetitive unit(s) consisting of module A and repetitive unit(s) consisting of module A
  • modules A, C, Q, K, sp, S, R, X or Y or variants thereof can also be combined with each other in any combination and in any number of each, i.e. module (repetitive unit) A can be combined with module (repetitive unit) Q (i.e. combination AQ), module (repetitive unit) C can be combined with module (repetitive unit) Q (i.e.
  • module (repetitive unit) Q can be combined with module (repetitive unit) A and with module (repetitive unit) Q (i.e. combination QAQ), module (repetitive unit) A can be combined with module (repetitive unit) A and with module (repetitive unit) Q (i.e. combination AAQ), etc., under the proviso that the silk polypeptide used in the method of the present invention comprises or consists of at least two repetitive units which are identical.
  • the silk polypeptide used in the method of the present invention can comprise or consist of A n , (AA) n , (AQ) n , (QA) n , Q n , (QQ) friendship, (QAQ)n, (AQA) n , C n , (CC) n , (CCC) discipline, (CQ) n , (QC) n , (QCQ) n , (CQC) n , (AA) n Q n ,
  • n is at least 2, preferably 4, 8, 9, 10, 12, 16, 20, 24, or 32.
  • the silk polypeptide consists of (AQ) 12
  • module (repetitive unit) A is 12 times present and module (repetitive unit) Q is also 12 times present in the silk polypeptide and that, thus, the silk polypeptide consists of 24 modules (repetitive units).
  • the arrangement of the modules (repeat units) of a silk polypeptide consisting of (AQ) 12 is as follows: AQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQAQ.
  • the silk polypeptide of the modules (repeat units) of a silk polypeptide consists of (QAQ) 8
  • module (repeat unit) A is 8 times present
  • module (repetitive unit) Q is 16 times present in the silk polypeptide and that, thus, the silk polypeptide consists of 24 modules (repetitive units).
  • the arrangement of the modules (repeat units) of a silk polypeptide consisting of (QAQ) 8 is as follows:
  • the silk polypeptide used in the method of the present invention can also comprise or consist of (A*Q) n , (AQ*) n , (A*Q*) friendship, (Q*A) n , (QA*) n , (Q*A*) n , (QAQ*) n , (Q*AQ) n , (QA*Q*) n , (Q*A*Q) usually, (Q*AQ*) n , (Q*A*Q*) n , (AQA*) n , (AQ*A) n , (AQ*A) n , (AQ*A) n , (AQ*A) n , (AQ*A*) n , (AQ*A*) n , (AQ*A*) n , (AQ*A*) n , (AQ*A*) n , (AQ*A*) n , (AQ*A*) intimate, (A*Q*A) n ,
  • modules (repetitive units) are directly combined or concatenated with each other or may mean in the context of the present invention that the modules (repetitive units) are combined or concatenated with each other via one or more spacer amino acids.
  • the modules (repetitive units) comprised in the silk polypeptide are directly combined or concatenated with each other.
  • the modules (repetitive units) comprised in the silk polypeptide are combined or concatenated with each other via 1 to 25 or 1 to 20 spacer amino acids, more preferably via 1 to 15 or 1 to 10 spacer amino acids, and most preferably, via 1 to 5 spacer amino acids, i.e. via 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 spacer amino acids.
  • Said spacer amino acids may be any amino acids naturally occurring in proteins.
  • said spacer amino acid is not proline. It is preferred that the spacer amino acid(s) contain(s) charged groups.
  • the spacer amino acid(s) containing charged groups is (are) independently selected from the group consisting of aspartate, glutamate, histidine, and lysine.
  • Said spacer amino acids should be amino acids which do not negatively affect the ability of a silk polypeptide to coat, preferably to uniformly coat, the inert or naturally occurring material, such as kevlar or wool.
  • said spacer amino acids should be amino acids which do not cause steric hindrance, e.g. amino acids having a small size such as lysine and cysteine.
  • the silk polypeptide comprises modules which are directly combined with each other and modules which are combined with each other via 1 to 25 or 1 to 20 spacer amino acids, more preferably via 1 to 15 or 1 to 10 spacer amino acids, and most preferably, via 1 to 5 spacer amino acids, i.e. via 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 spacer amino acids.
  • a module A, C, Q, K, sp, S, R, X or Y variant differs from the reference (wild- type) module A, C, Q, K, sp, S, R, X or Y from which it is derived by up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acid changes in the amino acid sequence (i.e. substitutions, additions, insertions, deletions, N-terminal truncations and/or C-terminal truncations).
  • Such a module variant can alternatively or additionally be characterised by a certain degree of sequence identity to the reference (wild-type) module from which it is derived.
  • a module A, C, Q, K, sp, S, R, X or Y variant has a sequence identity of at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or even 99.9% to the respective reference (wild- type) modul A, C, Q, K, sp, S, R, X or Y.
  • sequence identity is over a continuous stretch of at least 10, 15, 18, 20, 24, 27, 28, 30, 34, 35, or more amino acids, preferably over the whole length of the respective reference (wild-type) module A, C, Q, K, sp, S, R, X or Y.
  • sequence identity is at least 80% over the whole length, is at least 85% over the whole length, is at least 90% over the whole length, is at least 95% over the whole length, is at least 98% over the whole length, or is at least 99% over the whole length of the respective reference (wild-type) module A, C, Q, K, sp, S, R, X or Y.
  • sequence identity is at least 80% over a continuous stretch of at least 10, 15, 18, 20, 24, 28, or 30 amino acids, is at least 85% over a continuous stretch of at least 10, 15, 18, 20, 24, 28, or 30 amino acids, is at least 90% over a continuous stretch of at least 10, 15, 18, 20, 24, 28, or 30 amino acids, is at least 95% over a continuous stretch of at least 10, 15, 18, 20, 24, 28, or 30 amino acids, is at least 98% over a continuous stretch of at least 10, 15, 18, 20, 24, 28, or 30 amino acids, or is at least 99% over a continuous stretch of at least 10, 15, 18, 20, 24, 28, or 30 amino acids of the respective reference (wild-type) module A, C, Q, , sp, S, R, X or Y.
  • a fragment (or deletion variant) of module A, C, Q, , sp, S, R, X or Y has preferably a deletion of up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, or 15 amino acids at its N-terminus and/or at its C-terminus.
  • the deletion can also be internally.
  • the module A, C, Q, K, sp, S, R, X or Y variant or fragment is only regarded as a module A, C, Q, , sp, S, R, X or Y variant or fragment within the context of the present invention, if the modifications with respect to the amino acid sequence on which the variant or fragment is based do not negatively affect the ability of a silk polypeptide to form a fibre.
  • the skilled person can visually assess whether a fibre is still formed.
  • the smooth and homogenous appearance of said fibre can be controlled using el ectroni c-microscopy .
  • the repetitive units are independently selected from module A c (SEQ ID NO: 29), module A K (SEQ ID NO: 30), module C c (SEQ ID NO: 31), module C K1 (SEQ ID NO: 32), module C 2 (SEQ ID NO: 33) or module C KC (SEQ ID NO: 34).
  • C ICC (SEQ ID NO: 34) are variants of the module A which is based on the amino acid sequence of ADF-3 of the spider Araneus diadematus and of module C which is based on the amino acid sequence of ADF-4 of the spider Araneus diadematus (WO 2007/025719).
  • module A c the amino acid S (serine) at position 21 has been replaced by the amino acid C (cysteine)
  • module A K SEQ ID NO: 30
  • the amino acid S at position 21 has been replaced by the amino acid K (lysine)
  • module C SEQ ID NO: 31
  • the amino acid S at position 25 has been replaced by the amino acid 25 by C
  • module C K1 SEQ ID NO: 32
  • the amino acid S at position 25 has been replaced by the amino acid K
  • module C ⁇ the amino acid E (glutamate) at position 20 has been replaced by the amino acid K
  • module C KC SEQ ID NO: 34
  • the amino acid E at position 20 has been replaced by the amino acid K and the amino acid S at position 25 has been replaced by the amino acid C (WO 2007/025719).
  • the repetitive units in the silk polypeptide used in the method of the present invention consists of module A c : GPYGPGASAAAAAAGGYGPGCGQQ (SEQ ID NO: 29), module A K : GPYGPGASAAAAAAGGYGPGKGQQ (SEQ ID NO: 30), module C c : GSSAAAAAAAASGPGGYGPENQGPCGPGGYGPGGP (SEQ ID NO: 31), module C I : GSSAAAAAAAASGPGGYGPENQGPKGPGGYGPGGP (SEQ ID NO: 32), module C 1 ": GSSAAAAAAAAAASGPGGYGPKNQGPSGPGGYGPGGP (SEQ ID NO: 33), or module C C : GSSAAAAAAAASGPGGYGPKNQGPCGPGGYG PGGP (SEQ ID NO: 34).
  • the silk polypeptide used in the method of the present invention comprises, essentially consists of, or consists of between 2 to 80 repetitive units, between 3 to 80 repetitive units, or between 4 to 60 repetitive units, preferably between 8 to 48 repetitive units, or between 10 to 40 repetitive units and most preferably between 16 to 32 repetitive units, i.e. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,
  • the silk polypeptide used in the method of the present invention can comprises or consists of the modules r
  • the silk polypeptide used in the method of the present invention can also comprise or consist of the modules (A K ) n , (C c ) friendship, (C K1 ) n , (C K2 ) tradition, (C KC ) resort, (A K A c ) n , (C c C c ) n , (C K1 C K2 ) n , (C 2 C K, ) n , (C K1 C 2 C KI ) n , (C K2 C K1 C K2 ) n , (C ⁇ 'C ⁇ C ⁇ n , (C ⁇ CW ⁇ ),, or (C KC C K2 C K1 ) n , wherein n is at least 2, preferably 4, 5, 6, 7, 8, 10, 12,
  • the silk polypeptide used in the method of the present invention comprises, essentially consists of, or consists of between 2 to 80 repetitive units, between 3 to 80 repetitive units or between 4 to 60 repetitive units, preferably between 8 to 48 repetitive units, or between 10 to 40 repetitive units and most preferably between 16 to 32 repetitive units, i.e.
  • the modules A K , C c , C K1 , C 2 and C KC can also be combined with the modules A, C, Q, K, sp, S, R, X or Y, i.e. module (repetitive unit)
  • a K can be combined with module (repetitive unit) C (i.e. combination A K C), or module (repetitive unit) C c can be combined with module (repetitive unit) C (i.e. combination C C C), etc., under the proviso that the silk polypeptide used in the method of the present invention comprises or consists of at least two repetitive units which are identical.
  • the silk polypeptide used in the method of the present invention comprises or consists of the modules C 16 C C , C C C 16 , C 8 C C C 8 , C 8 C C 8 , C C 8 C 8 , C 4 C C 8 C 4 , C C 4 C 8 C C 4 , C C (AQ) 24 , or (AQ) 24 C C .
  • the silk polypeptide used in the method of the present invention can further comprise at least one non-repetitive (NR) unit, i.e. 1 , 2, 3, 4, 5, 6, or more NR units, preferably one NR unit.
  • NR non-repetitive
  • the term "non-repetitive (NR) unit” refers to a region of amino acids present in a naturally occurring silk polypeptide that displays no obvious repetition pattern (non -repetitive unit or NR unit).
  • the amino acid sequence of the non-repetitive unit corresponds to a non- repetitive amino acid sequence of naturally occurring dragline polypeptides, preferably of ADF-3 (SEQ ID NO: 1) or ADF-4 (SEQ ID NO: 2), or to an amino acid sequence substantially similar thereto.
  • the amino acid sequence of the non-repetitive unit corresponds to a non-repetitive carboxy terminal amino acid sequence of naturally occurring dragline polypeptides, preferably of ADF-3 (SEQ ID NO: 1) or ADF-4 (SEQ ID NO: 2), or to an amino acid sequence substantially similar thereto. More preferably, the amino acid sequence of the non-repetitive unit corresponds to a non-repetitive carboxy terminal amino acid sequence of ADF-3 (SEQ ID NO: 1) which comprises amino acids 513 through 636, or of ADF-4 (SEQ ID NO: 2) which comprises amino acids 302 through 410, or to an amino acid sequence substantially similar thereto.
  • substantially similar means a degree of amino acid identity of at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or even 99.9%, preferably over 20, 30, 40, 50, 60, 70, 80 or more amino acids, more preferably over the whole length of the respective reference non-repetitive (carboxy terminal) amino acid sequence of naturally occurring dragline polypeptides, preferably of ADF-3 (SEQ ID NO: 1) or ADF-4 (SEQ ID NO: 2) ⁇
  • non-repetitive unit having an amino acid sequence which is "substantially similar” to a corresponding non-repetitive (carboxy terminal) amino acid sequence within a naturally occurring dragline polypeptide (i.e. wild-type non-repetitive (carboxy terminal) unit), preferably within ADF-3 (SEQ ID NO: 1) or ADF-4 (SEQ ID NO: 2), is also similar with respect to its functional properties, e.g. a silk polypeptide comprising the "substantially similar non-repetitive unit” still has the ability to form a fibre.
  • a smooth and homogenous fibre from the silk polypeptide comprising the "substantially similar non-repetitive unit" at a speed of at least 0.1 cm/s, preferably 10 cm/s and more preferably 10 m/s.
  • the skilled person can visually assess whether a fibre is still formed.
  • the smooth and homogenous appearance of said fibre can be controlled using electronic-microscopy.
  • the non-repetitive (NR) unit is NR3 (SEQ ID NO: 41) or variants thereof, or NR4 (SEQ ID NO: 42) or variants thereof.
  • the NR3 (SEQ ID NO: 41) unit is based on the amino acid sequence of ADF-3 of the spider Araneus diadematus and the NR4 (SEQ ID NO: 42) unit is based on the amino acid sequence of ADF-4 of the spider Araneus diadematus (WO 2006/008163).
  • a NR3 or NR4 unit variant differs from the reference NR3 (SEQ ID NO: 41) or NR4 (SEQ ID NO: 42) unit from which it is derived by up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, or 30 amino acid changes in the amino acid sequence (i.e. exchanges, insertions, deletions, N-terminal truncations and/or C-terminal truncations).
  • Such a NR3 or NR4 unit variant can alternatively or additionally be characterised by a certain degree of sequence identity to the reference NR3 or NR4 unit from which it is derived.
  • a NR3 or NR4 unit variant has a sequence identity of at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or even 99.9% to the respective reference NR3 or NR4 unit.
  • the sequence identity is over a continuous stretch of at least 10, 20, 30, 40, 50, 60, 70, 80, 90, or more amino acids, preferably over the whole length of the respective reference NR3 or NR4 unit.
  • sequence identity is at least 80% over the whole length, is at least 85% over the whole length, is at least 90% over the whole length, is at least 95% over the whole length, is at least 98% over the whole length, or is at least 99% over the whole length of the respective reference NR3 or NR4 unit.
  • sequence identity is at least 80% over a continuous stretch of at least 20, 30, 40, 50, 60, 70, or 80 amino acids, is at least 85% over a continuous stretch of at least 20, 30, 40, 50, 60, 70, or 80 amino acids, is at least 90% over a continuous stretch of at least 20, 30, 40, 50, 60, 70, or 80 amino acids, is at least 95% over a continuous stretch of at least 20, 30, 40, 50, 60, 70, or 80 amino acids, is at least 98% over a continuous stretch of at least 20, 30, 40, 50, 60, 70, or 80 amino acids, or is at least 99% over a continuous stretch of at least 20, 30, 40, 50, 60, 70, or 80 amino acids of the respective reference NR3 or NR4 unit.
  • a fragment (or deletion variant) of a NR3 or NR4 unit has preferably a deletion of up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, or 60 amino acids at its N-terminus and/or at its C-terminus.
  • the deletion can also be internally.
  • the NR3 or NR4 unit variant or fragment is only regarded as a NR3 or NR4 unit variant or fragment within the context of the present invention, if the modifications with respect to the amino acid sequence on which the variant or fragment is based do not negatively affect the ability of a silk polypeptide to form a fibre.
  • the skilled person can visually assess whether a fibre is still formed.
  • the smooth and homogenous appearance of said fibre can be controlled using electronic-microscopy.
  • NR3 or NR4 unit variant or fragment still enables the polymerization and/or increases the solubility of a silk polypeptide wherein it is comprised.
  • the skilled person can readily assess whether a silk polypeptide comprising a NR3 or NR4 unit variant or fragment has the above mentioned functional properties like a silk polypeptide comprising the respective reference NR3 or NR4 unit. Suitable assays are well known to the person skilled in the art.
  • the polymerization of silk polypeptides comprising a NR3 or NR4 unit variant or fragment and the polymerization of silk polypeptides comprising the respective reference NR3 or NR4 unit can easily be visualized via native gel electrophoresis.
  • solubility of a silk polypeptide comprising a NR3 or NR4 unit variant or fragment and the solubility of a silk polypeptide comprising the respective reference NR3 or NR4 unit can simply be tested via saturation of said silk polypeptides in an aqueous solution. The results can finally be compared with each other.
  • the silk polypeptide used in the method of the present invention is selected from the group consisting of ADF-3 (SEQ ID NO: 1) or variants thereof, ADF- 4 (SEQ ID NO: 2) or variants thereof, MaSp I (SEQ ID NO: 43) or variants thereof, MaSp II (SEQ ID NO: 44) or variants thereof, (C) m , (C) m NR z , NR z (C) m , (AQ) n , (AQ) n NR z , NR z (AQ) n , (QAQ) 0 , NR Z (QAQ) 0 , (QAQ) 0 NR 2 , Y p , X p , and p , wherein m is an integer of 8 to 48 (i.e.
  • n is an integer of 4 to 24 (i.e. 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24)
  • o is an integer of 2 to 20 (i.e. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20)
  • p is an integer of 8 to 16 (i.e. 8, 9, 10, 11, 12, 13, 14, 15, or 16)
  • z is an integer of 1 to 3 (i.e. 1, 2, or 3), preferably 1, and NR stands for a non- repetitive unit.
  • n NR z a "n" number of A and Q module combinations, namely 6 to 24 A and Q module combinations, wherein module A is represented by the amino acid sequence according to SEQ ID NO: 20 and module Q is represented by the amino acid sequence according to SEQ ID NO: 22, are combined with each other, and wherein said A and Q module combinations are further combined with a "z" number of non-repetitive (NR) units, namely 1 to 3 non-repetitive (NR) units, e.g. the non- repetitive (NR) units NR3 represented by the amino acid sequence according to SEQ ID NO: 41 or NR4 represented by the amino acid sequence according to SEQ ID NO: 42,
  • NR non-repetitive
  • NR non-repetitive
  • z in (C) m NR z or NR z (C) m is 1 and m is an integer of 8 to 48 (i.e. 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48), z in (C) n NR z or NR z (C) m is 2 and m is an integer of 8 to 48 (i.e.
  • NR Z (QAQ) 0 or (QAQ) 0 NR Z is 1 and o is an integer of 2 to 20 (i.e. 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, or 20), z in NR z (QAQ)o or (QAQ) 0 NR Z is 2 and o is an integer of 2 to 20 (i.e. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20), or z in NR z (QAQ)o or (QAQ) 0 NR Z is 3 and o is an integer of 2 to 20 (i.e. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20), wherein NR stands for a non-repetitive unit, preferably NR3 or NR4.
  • the silk polypeptide used in the method of the present invention is C 16 NR3, C 32 NR3, C 16 NR4, C 32 NR4, (AQ) 12 , (AQ) 24 , (AQ) 12 NR3, (AQ) 24 NR3, (AQ) 12 NR4, (AQ) 24 NR4, C 16 , C 32 , Y 8 , Y 16 , X g , X 16 , 8 , or i 6 .
  • An ADF-3, ADF-4, MaSp I or MaSp II variant differs from the reference (wild- type) ADF-3 (SEQ ID NO: 1), ADF-4 (SEQ ID NO: 2), MaSp I (SEQ ID NO: 43) or MaSp II (SEQ ID NO: 44) polypeptide from which it is derived by up to 150 (up to 1, 2, 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, or 150) amino acid changes in the amino acid sequence (i.e.
  • Such a variant can alternatively or additionally be characterised by a certain degree of sequence identity to the reference (wild-type) polypeptide from which it is derived.
  • an ADF-3, ADF-4, MaSp I or MaSp II variant has a sequence identity of at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or even 99.9% to the respective reference (wild-type) ADF-3, ADF-4, MaSp I or MaSp II polypeptide.
  • sequence identity is over a continuous stretch of at least 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 120, 150, 180, 200, 250, 300, 350, 400, or more amino acids, preferably over the whole length of the respective reference (wild-type) ADF-3, ADF-4, MaSp I or MaSp II polypeptide.
  • sequence identity is at least 80% over the whole length, is at least 85% over the whole length, is at least 90% over the whole length, is at least 95% over the whole length, is at least 98%> over the whole length, or is at least 99% over the whole length of the respective reference (wild-type) ADF-3, ADF- 4, MaSp I or MaSp II polypeptide.
  • sequence identity is at least 80% over a continuous stretch of at least 20, 30, 50, 100, 150, 200, 250, or 300 amino acids, is at least 85% over a continuous stretch of at least 20, 30, 50, 100, 150, 200, 250, or 300 amino acids, is at least 90% over a continuous stretch of at least 20, 30, 50, 100, 150, 200, 250, or 300 amino acids, is at least 95% over a continuous stretch of at least 20, 30, 50, 100, 150, 200, 250, or 300 amino acids, is at least 98% over a continuous stretch of at least 20, 30, 50, 100, 150, 200, 250, or 300 amino acids, or is at least 99% over a continuous stretch of at least 20, 30, 50, 100, 150, 150,
  • a fragment (or deletion variant) of the ADF-3 (SEQ ID NO: 1) polypeptide has preferably a deletion of up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 120, 150, 170, 200, 220, 250, 270, 300, 320, 350,
  • the deletion can also be internally.
  • a fragment (or deletion variant) of the ADF-4 (SEQ ID NO: 2) polypeptide has preferably a deletion of up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 120, 150, 170, 200, 220, 250, 270, 300, 320, 330,
  • the deletion can also be internally.
  • a fragment (or deletion variant) of the MaSp I (SEQ ID NO: 43) polypeptide has preferably a deletion of up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550,
  • a fragment (or deletion variant) of the MaSp II (SEQ ID NO: 44) polypeptide has preferably a deletion of up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 520,
  • deletion can also be internally.
  • the ADF-3, ADF-4, MaSp I or MaSp II variant or fragment is only regarded as an ADF-3, ADF-4, MaSp I or MaSp II variant or fragment within the context of the present invention, if the modifications with respect to the amino acid sequence on which the variant or fragment is based do not negatively affect the ability of a silk polypeptide to form a fibre.
  • the skilled person can visually assess whether a fibre is still formed.
  • the smooth and homogenous appearance of said fibre can be controlled using electronic-microscopy.
  • the method is carried out at temperatures of between 5°C and 60°C, more preferably at temperatures of between 10°C and 50°C and most preferably, at temperatures of between 20°C and 40°C.
  • the method can be carried out at a temperature of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60°C.
  • the method is carried out at a pressure of between 10 kPa and 1000 kPa, more preferably between 40 kPa and 500 kPa and most preferably between 50 kPa and 150 kPa.
  • the method can be carried out at 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 950, 1000 kPa. It is particularly preferred that the method is carried out at standard conditions for temperature and pressure, i.e. at 20°C and 101.325 kPa.
  • the spinning solution provided in step (a) comprises more than one polymer, preferably 2, 3, or 4 polymers, e.g. polypeptides such as silk polypeptides, casein, zein or BSA.
  • the spinning solution provided in step (a) can comprise (i) a silk polypeptide and casein, (ii) a silk polypeptide and zein, (iii) a silk polypeptide and BSA, (iv) a silk polypeptide, zein, and casein, (v) a silk polypeptide, casein and BSA, (vi) a silk polypeptide, BSA, casein and zein, (vii) zein and casein, (viii) zein and BSA, (ix) casein and BSA, or (x) zein and casein and BSA.
  • the polypeptides can be labeled, for example, with enzymatic labels, biotin, radioactive or fluorescent labels, e.g. fluor
  • the spinning solution provided in step (a) comprises more than one type of silk polypeptide.
  • the spinning solution provided in step (a) of the method of the present invention comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 different types of silk polypeptides, most preferably 2 different types of silk polypeptides.
  • the spinning solution can comprise dragline spider silk polypeptides, which differ from each other with respect to their amino acid sequence.
  • the solution provided in the method of the present invention can also comprise dragline spider silk and flagelliform spider silk polypeptides which differ from each other with respect to their natural origin.
  • the dragline spider silk polypeptide is from the major ampullate gland, while the flagelliform polypeptide is from the flagelliform gland.
  • the concentration of the polymer is preferably in the range of 0.15 mg/ml to 500 mg/ml, more preferably in the range of 0.15 mg/ml to 99.9 mg/ml, in the range of 0.15 mg/ml to 90 mg/ml, in the range of 0.15 mg/ml to 80 mg/ml, in the range of 0.15 mg/ml to 70 mg/ml, in the range of 0.15 mg/ml to 60 mg/ml, or in the range of 0.15 mg/ml to 50 mg/ml, most preferably in the range of 0.5 mg/ml to 50 mg/ml, in the range of 0.5 mg/ml to 40 mg/ml, in the range of 0.5 mg/ml to 30 mg/ml or in the range of 1 mg/ml to 20 mg/ml. It is particularly preferred that the concentration of the polymer, e.g. silk polypeptide, BSA, zein or casein.
  • the concentration of the polymer is (at least) 0.15 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4 mg/ml, 0.5 mg/ml, 0.6 mg/ml, 0.7 mg/ml, 0.8 mg/ml, 0.9 mg/ml, 1 mg/ml, 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, 10 mg/ml, 11 mg/ml, 12 mg/ml, 13 mg/ml, 14mg/ml, 15 mg/ml, 16 mg/ml, 17 mg/ml, 18 mg/ml, 19 mg/ml, 20 mg ml, 21 mg/ml, 22 mg/ml, 23 mg/ml, 24 mg/ml, 25 mg/ml, 26 mg m
  • the method of the present invention requires only very low concentrations of polymers, e.g. silk polypeptides, beginning at concentrations as low as 0.15 mg/ml and, preferably, ending at concentrations as not higher as 90 mg/ml, more preferably as not higher as 50 mg/ml and most preferably as not higher as 20 mg/ml.
  • This has the advantage that the shear viscosity of the spinning solution used in the method of the present invention is very low, which, in turn, improves the processability of said spinning solution.
  • the viscosity increasing compound is not limited in its structure as long as it increases the elongational viscosity of the spinning solution.
  • the compound that increases elongational viscosity of the solution is a linear or branched polymer with a molecular weight of at least 20 kDa, e.g.
  • kDa 25 kDa, 30 kDa, 40 kDa, 50 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa, 100 kDa, 200 kDa, 300 kDa, 400 kDa, 500 kDa, 600 kDa, 700 kDa, 800 kDa, 900 kDa, 1 MDa, 1.5 MDa, 2 MDa, 2.5 MDa, 3 MDa, 3.5 MDa, 4 MDa, 4.5 MDa, or 5 MDa.
  • the linear or branched polymer has a molecular weight of between 20 kDa to 5 MDa or of between 50 kDa to 4 MDa, more preferably of between 100 kDa to 2 MDa or of between 100 kDa to 1 MDa.
  • the compound that increases elongational viscosity of the solution is a linear or branched polymer with a molecular weight of at least 500 kDa, e.g. 550 kDa, 600 kDa, 650 kDa, 700 kDa, 750 kDa, 800 kDa, 850 kDa, 900 kDa, 950 kDa, 1 MDa, 1.5 MDa, 2 MDa, 2.5 MDa, 3 MDa, 3.5 MDa, 4 MDa, 4.5 MDa, or 5 MDa.
  • 500 kDa e.g. 550 kDa, 600 kDa, 650 kDa, 700 kDa, 750 kDa, 800 kDa, 850 kDa, 900 kDa, 950 kDa, 1 MDa, 1.5 MDa, 2 MDa, 2.5 MDa, 3 MDa, 3.5 MDa, 4 MDa, 4.5 MDa, or 5
  • the linear or branched polymer has a molecular weight of between 500 kDa to 5 MDa or of between 900 kDa to 5 MDa, most preferably of between 1 MDa to 4 MDa or of between 1 MDa to 3 MDa.
  • the compound that increases elongational viscosity of the solution is selected from the group consisting of polyacrylamide (PAA), polyethylene (PE), polyethylene glycol (PEG), polysaccharides, dextrans, polyvinyl-alcohols, nucleic acids, and mixtures thereof.
  • PAA polyacrylamide
  • PE polyethylene
  • PEG polyethylene glycol
  • dextrane is dextrane-sulfate.
  • the compound that increases elongational viscosity of the solution is polyethylene glycol (PEG) with a molecular weight of at least 20 kDa, e.g. 25 kDa, 30 kDa, 40 kDa, 50 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa, 100 kDa, 200 kDa, 300 kDa, 400 kDa, 500 kDa, 600 kDa, 700 kDa, 800 kDa, 900 kDa, 1 MDa, 1.5 MDa, 2 MDa, 2.5 MDa, 3 MDa, 3.5 MDa, 4 MDa, 4.5 MDa, or 5 MDa.
  • PEG polyethylene glycol
  • polyethylene glycol has a molecular weight of between 20kDa to 5 MDa or of between 50 kDa to 4 MDa, more preferably of between 100 kDa to 2 MDa or of between 100 kDa to 1 MDa.
  • the compound that increases elongational viscosity of the solution is polyethylene glycol (PEG) with a molecular weight of at least 500 kDa, e.g. 550 kDa, 600 kDa, 650 kDa, 700 kDa, 750 kDa, 800 kDa, 850 kDa, 900 kDa, 950 kDa, 1 MDa, 1.5 MDa, 2 MDa, 2.5 MDa, 3 MDa, 3.5 MDa, 4 MDa, 4.5 MDa, or 5 MDa.
  • PEG polyethylene glycol
  • polyethylene glycol has a molecular weight of between 500 kDa to 5 MDa or of between 900 kDa to 5 MDa, most preferably of between 1 MDa to 4 MDa or of between 1 MDa to 3 MDa.
  • the concentration of the compound that increases elongational viscosity of the solution is preferably in the range of 0.1 wt%/vol to 5 wt%/vol, more preferably in the range of 0.5 wt%/vol to 4 wt%/vol, and most preferably in the range of 1 wt%/vol to 3 wt%/vol.
  • the concentration of the compound that increases elongational viscosity of the solution is 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5 4.6, 4.7, 4.8, 4.9, or 5 wt%/vol.
  • the inventors of the present invention surprisingly found that already this low concentration of the compound that increases elongational viscosity of the solution is sufficient for spinning fibres with the method provided herein.
  • the low concentration of said compound increases on the one hand side the elongational viscosity of the solution, but on the other hand side does not substantially increase the shear viscosity of the solution so that the generation of fibres is improved.
  • the spinning solution used in the method of the present invention is mainly composed of a polymer, such as a silk polypeptide, zein or casein, i.e. is relatively pure.
  • the method of the present invention is cost efficient and ecologically friendly.
  • the ratio by weight% of the silk polypeptide to the elongational viscosity enhancer is preferably between 0.1 to 5, 0.2 to 5, 0.3 to 5, or 0.4 to 5, more preferably between 0.4 to 2, most preferable between 0.8 to 1.5.
  • the amount of elongational viscosity enhancer is chosen in such that the ratio by weight% of the silk polypeptide to the elongational viscosity enhancer is preferably between 0.1 to 10, more preferably between 0.2 to 5, more preferably between 0.4 to 2, most preferable between 0.8 to 1.5.
  • the Ohnesorge number of an C 16 spinning solution can be increased sufficiently in order to allow improved fibre formation at the air/liquid interface.
  • the solvent can be any solution in which the polymer, e.g. the polypeptide such as the spider silk polypeptide, zein, BSA or casein, and the elongational viscosity enhancing polymer are soluble.
  • the solvent is selected from the group consisting of a polar solvent, preferably water, an aqueous buffer, an alcohol, preferably isopropanol, Hexafluoroisopropanol, glycerol ethanol (EtOH), propanol, butanol, octanol, or acetone; a non-polar solvent, preferably hexane, dodecane or oil, other organic solvents, preferably ethylacetate; and mixtures thereof.
  • the solvent can also be a mixture of a polar solvent, e.g. water, and a non-polar solvent, e.g. hexane.
  • the pH of the solvent is between 4 and 10.
  • fibre formation can be facilitated with the method of the present invention.
  • the drawing of the fibre is initiated by contacting an air/liquid or liquid/liquid interface of the solution with a drawing tool and withdrawing the drawing tool from said interface.
  • air as used herein preferably means the mixture of gases naturally occurring in the earth atmosphere.
  • the solvent of the spinning solution is a polar solvent the other liquid is preferably a non-polar.
  • the solvent of the spinning solution is a non-polar solvent preferably the other liquid is polar.
  • a liquid/liquid interface may also be formed at least temporarily when two miscible liquids are brought into contact, e.g.
  • the one liquid stream is enclosing a stream of the second liquid.
  • a drawing tool e.g. a pipette tip
  • the withdrawing of the pipette leads to an orientation of the polymers in the solvent with its head or tail towards the drawing tool, whereby the polymers align side-by-side and increasingly associate with each other.
  • the polymers transition from an ordered solution or liquid crystal type state to a semi solid or solid state.
  • drawing is used in this context not only refer to the initiation of this alignment process but also to refer to continued withdrawing of aligned or partially aligned polymer molecules from the spinning solution.
  • the drawing tool no longer contacts the spinning solution but the "drawing" of further polymer molecules from the solution into a "detached solution", i.e. a solution comprising aligned polymers no longer in contact with the body of the spinning solution.
  • drawing means drawing as a process of fibre formation, which can also include stretching or hardening.
  • drawing the fibre from the solution is not a process restricted to pulling a fibre from a detached solution with a drawing tool like, e.g.
  • the process can also comprise any appropriate automated system, for example, a pipe through which the solution is driven in a replenishing flow, with a nozzle at the end where the spinning solution extrudes and is being guided away by a drawing tool, thereby continuously drawing the fibre from the extruded spinning solution.
  • the drawing tool can be any kind of instrument with a tip appropriate for drawing a fibre and of an appropriate material, e.g. capable of providing an attachment point to the polymers in the spinning solution. Examples include, but are not limited to pipette tips, needles or thin bars made of plastic, metal or glass.
  • the speed of fibre-drawing is not limited, as long as it is high enough for drawing a thin fluid filament, which exists long enough to allow evaporation of the solvent and the formation of a fibre, preferably a thin, light and long fibre.
  • drawing is carried out by touching the solution with a drawing tool and the drawing of a fibre from the surface.
  • drawing also refers to the withdrawing of the fibre from the solution, preferably at a constant speed. The ideal speed depends on the viscosity of the spinning solution, i.e. on type and concentration of the polymers used, and has to be determined by experiment.
  • the drawing is carried out at speeds of at least 0.1 cm/s, preferably between 0.1 cm/s and 15 m/s, between 1 cm/s and 5 m/s, between 3 cm/s and 1 m/s, between 5 cm/s and 50 cm/s, between 5 cm/s and 15 cm/s, and more preferably at 10 cm/s.
  • the drawing is carried out at speeds of at least 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 30, 40, 50, 60, 70, 80, 90 cm/s, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 m/s.
  • the spinning solution is extruded to form a fibre.
  • extruded in this context means the application of pressure to the spinning solution to force it through an opening, e.g. a nozzle.
  • extruded spinning solution does not solidify entirely. Rather the dissolved polymers that are comprised in the extruded spinning solution associate to form a fibre, which is typically smaller in diameter than the opening through which the spinning solution is extruded, while the remaining solvent is separated from the fibre thus formed.
  • the fibre initially formed is attached to a fibre recovery device, e.g. a cylinder, spool or bobbin, onto which the fibre is continuously wound.
  • a fibre recovery device e.g. a cylinder, spool or bobbin
  • the fibre is continuously wound.
  • a pulling force exerted onto the extruded fibre i.e. the fibre in some embodiments may be considered to be "drawn" from to extruded solution.
  • the shear viscosity of artificial spinning dopes can be increased simultaneously to the elongational viscosity to an amount not detrimental to fluid processing by, for example, adding Newtonian viscosity enhancers.
  • Spinnability thereby means the suitability of a spinning solution for fibre drawing.
  • the spinning solution further comprises a compound that increases Newtonian viscosity, preferably monosaccharides, e.g. glucose, galactose, or fructose; disaccharides, e.g. lactose, or sucrose; polysaccharides, e.g. glycogen; or polyols, e.g. inositiol, sorbitol, glycerol, or polyglycols; or mixtures thereof.
  • a compound that increases Newtonian viscosity preferably monosaccharides, e.g. glucose, galactose, or fructose; disaccharides, e.g. lactose, or sucrose; polysaccharides, e.g. glycogen; or polyols, e.g. inositiol, sorbitol, glycerol, or polyglycols; or mixtures thereof.
  • the spinning solution further comprises a particulate matter dispersed therein.
  • this particulate matter is embedded in the fibre in a relatively uniform manner.
  • the particulate matter is not limited in its composition as long as it is suitable for being incorporated into a fibre. It is preferred that the particulate matter is selected from the group consisting of beads, electricity conducting metals (including gold, copper), crystallites, protein/protein-complexes, enzymes (e.g. green fluorescent protein (GFP) or ⁇ -galactosidase) and cells (e.g. human osteoblast cells or human tendon precursor cells or any pluripotent cells or stem cells).
  • the cells are selected from enzyme or hormone secreting cells, e.g. Langerhans islet cells.
  • Preferred crystallites are dyes which are not soluble in water, fluorescent dyes (e.g. Sudan red) or azo dyes (e.g. Nile red).
  • the concentration of the particulate matter in the spinning solution is not limited.
  • the particulate matter is present in the spinning solution in an amount in the range of 0.01 wt%/vol to 10 wt%/vol, preferably 0.5 wt%/vol to 1 wt%/vol.
  • the particulate matter can be present in the spinning solution in an amount of 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0 wt%/vol.
  • the spinning solution further comprises a compound that increases Newtonian viscosity, e.g. polyglycols, or a particulate matter, e.g. beads.
  • the spinning solution further comprises both a compound that increases Newtonian viscosity, e.g. polyglycols, and a particulate matter, e.g. beads.
  • the fibre can be treated after formation by curing in a curing solution or cross- linking.
  • the method of the present invention comprises subsequent to step (b) a step (c) of curing the fibre in a curing solution or cross-linking the fibre.
  • Cross-linking can comprise linking other compounds to the fibre or linking identical or different fibres to each other, whereby, for example, multi-fibre-structures such as multi-layer fibres or core-shell structures can be produced.
  • cross-linking refers to a process of chemically joining two molecules by a covalent bond.
  • Cross-linking or coupling reagents contain reactive ends to specific functional groups (primary amines, sulfhydryl, etc.) on polypeptides or on other molecules.
  • the cross-linking or coupling reagent used in the method of the present invention is l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) or Glutaraldehyde.
  • the fibre is coiled while continuously drawn. This allows the production and storage of long fibres.
  • the present invention relates to a spinning solution for carrying out the method of the present invention, comprising, essentially consisting of or consisting of (i) a polymer that can be brought in to an aqueous solution in a concentration of at least 0.15 mg ml, (ii) a compound that increases elongational viscosity of the spinning solution and (iii) a solvent.
  • the polymer is a silk polypeptide as defined above.
  • the concentration of the polymer, e.g. silk polypeptide is at least 0.15 mg/ml and not more than 60 mg/ml, more preferably at least 0.15 mg/ml and not more than 50 mg/ml, and most preferably at least 0.15 mg/ml and not more than 20 mg/ml.
  • the concentration of the compound that increases elongational viscosity of the spinning solution is in the range of 0.1 wt%/vol to 5 wt%/vol, more preferably in the range of 0.5 wt%/vol to 4 wt%/vol, and most preferably in the range of 1 wt%/vol to 3 wt%/vol.
  • the concentration of the polymer is at least 0.15 mg/ml and not more than 60 mg/ml, more preferably at least 0.15 mg/ml and not more than 50 mg/ml, and most preferably at least 0.15 mg/ml and not more than 20 mg/ml and that the concentration of the compound that increases elongational viscosity of the spinning solution is in the range of 0.1 wt%/vol to 5 wt%/vol, more preferably in the range of 0.5 wt%/vol to 4 wt%/vol, and most preferably in the range of 1 wt%/vol to 3 wt%/vol.
  • the present invention provides a fibre obtainable by the method of the first aspect.
  • the present invention provides products comprising the fibre of the second aspect, such as
  • the present invention provides the use of a fibre or thread comprising one or more cells to assay one or more cellular functions and/or to maintain said cells.
  • the cellular function is assayed prior, during and/or after exposing the one or more cells to a test compound or to a stimulus, preferably to a mechanical, thermal and/or electric stimulus.
  • Cell-containing fibres could also serve as scaffold material for tissue engineering and the growth or artificial organs.
  • the present invention improves the spinnability of a polymer solution, allowing for the production of a whole range of novel materials.
  • the present invention is particularly suited for the incorporation of living and non-living biological material, because the process can be carried out at appropriate conditions regarding temperature, pressure, solvent and chemicals used.
  • any enzymatic activity can in principle be incorporated into such fibres.
  • cells can be embedded in fibres as well, as has been shown with the incorporation of colloidal micron-size particles into fibres. Since the conditions favour cell viability, applications in bioengineering and tissue culture are possible. Cell- containing fibres could serve as scaffold material for tissue engineering and the growth of artificial organs. Also, encapsulating living cells allows subjecting those cells to differing environmental conditions, by pulling them through a bath with reactants, for instance. With such approaches, new tools for cell biology may be accessible. In this spirit, the invention can be used for serial and also high-throughput manipulation of cells on a string.
  • fibres containing cells secreting wound healing factors could be incorporated into or supplement bandages or xerogels, particularly in a wet wound healing context.
  • Conceivable are also uses in the production of skin grafts, replacement ligaments or surgical mesh.
  • fibres produced according to the invention can be used in a wide range of other commercial products, for example rope, netting, clothing fabric, construction material, tent fabric, backpacks and in many others which have strength, elasticity and low weight as desired characteristics.
  • Fig.: 1 shows fibres drawn from a spinning solution comprising 5 mg/ml FITC-Ci 6 and 0.5 wt%/vol PAA. A plain glass needle is pulled away from an aliquot of spinning solution (A), resulting in the formation of a thin fluid filament (B).
  • Fibres drawn at a speed of at least 10 cm/s appear smooth and homogenous.
  • Fig.: 2 shows fibres drawn from a spinning solution comprising approximately 10 7 /ml
  • Polystyrene beads (1 ⁇ diameter) dispersed in 5 mg/ml Cj 6 and 0.5 wt%/vol PAA. The beads are incorporated in the fibres and distributed fairly homogenously.
  • Fig.: 3 shows Atomic Force Microscopy Image of a fibre drawn from a spinning solution comprising Polystyrene beads (1 ⁇ diameter) dispersed in 5 mg/ml
  • Ci 6 and 0.5 wt%/vol PAA The beads are covered with fibre material and are tightly connected to the fibre.
  • Fig.: 4 shows zeine fibre drawn from a spinning solution comprising 300 mg/ml zeine and 0.5% wt/vol PAA in 63.75% ethanol.
  • the zeine fibre exhibited a smooth surface.
  • Fig. 1 A shows an aliquot of spinning solution consisting of C] 6 as the constituent polymer, PAA as the elongational viscosity enhancing polymer, and water as the solvent on a glass microscopy slide, in contact with a plain glass needle as the drawing tool. Pulling the needle, which is in contact with the spinning solution, away from the solution produces a thin fluid filament. This filament, if drawn quickly enough, exists long enough to allow evaporation of the solvent water and the formation of a thin and light fibre (Fig. 1 B).
  • the fibres contain protein, which can be shown by IR spectroscopy and by fluorescence microscopy when labelled protein is used.
  • Example 2 Fibres from a low concentration spider silk protein
  • Example 3 Fibres pulled by a falling steel bead
  • Dynabeads Polystyrene or PS beads
  • 100 ⁇ spinning solution 10 mg/ml Ci 6 , 1.0 wt%/vol PAA
  • a plain glass needle (5 cm long and 1 mm in diameter) was brought into contact with the spinning solution and then pulled away from the microscopy slide by hand at a speed of approximately 10 cm/s. This process resulted in a fibre containing PS beads being pulled from the spinning solution.
  • the beads were embedded in the fibre and distributed fairly homogenously (Fig. 2). Atomic Force Microscopy showed that the beads were covered with fibre material and were tightly connected to the fibre (Fig. 3).
  • Zeine is a storage protein from corn and has been used as a material for decades. It is available in reasonable purity in bulk quantities at low cost (Tsai 1979). 300 mg/ml zeine in 85% ethanol were mixed with 2% wt/vol PAA, with a final PAA concentration of 0.5% wt/vol. 100 ⁇ of this spinning solution were placed on a glass microscopy slide. A plain glass needle (5 cm long and 1 mm in diameter) was brought into contact with the spinning solution and then pulled away from the microscopy slide by hand at a speed of approximately 10 cm/s. This process resulted in a zeine fibre being pulled from the spinning solution (Fig. 4). Such zeine fibres were insoluble in water and had a smooth surface. They appeared to have good mechanical stability, but were rather brittle and not ductile. Zeine fibres could be turned more ductile in a formaldehyde bath.
  • Example 6 Incorporating of crystallites into fibres
  • Dyes (Sudan Red (Sigma 20161-8) or Nile Red (Fluka 72485) were added as a fine powder to the spinning solution (10 mg/ml Ci 6 , 1.0 wt%/vol PAA) in suspension. Both dyes have been incorporated into the fibres. Both dyes are hydrophobic and adhere strongly to the hydrophobic spider silk proteins.
  • Fluorescent proteins such as Green Fluorescent Protein (GFP) have been incorporated into the fibres (10 mg/ml Ci , 1.0 wt%/vol PAA). After incorporation bright fluorescence has been observed. This shows that the folding of the protein (GFP) in the fibers and therefore the protein itself is still intact.
  • GFP Green Fluorescent Protein
  • ⁇ -galactosidase has been incorporated into the fibers (10 mg/ml Ci 6 , 1.0 wt%/vol PAA).
  • o-nitrophenyl- -Dgalactopyranosid ONPG
  • the concentration of o- nitrophenol is determined to quantify the activity of ⁇ -galactosidase in the fibre.
  • the incorporation of enzymes can be used for any applications where enzymes that are large enough to be trapped in the fibre while substances can diffuse in and out are needed.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Textile Engineering (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Manufacturing & Machinery (AREA)
  • Insects & Arthropods (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Mechanical Engineering (AREA)
  • Peptides Or Proteins (AREA)
  • Materials For Medical Uses (AREA)
  • Artificial Filaments (AREA)

Abstract

La présente invention concerne un procédé de filage d'une fibre contenant un polymère polypeptide. L'invention concerne en outre une fibre polymère pouvant être obtenue par ledit procédé et les utilisations associées. L'invention concerne également des produits comprenant ladite fibre polymère.
PCT/EP2011/001307 2010-03-17 2011-03-16 Procédé de fabrication de fibres contenant un polymère WO2011113592A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
AU2011229482A AU2011229482B2 (en) 2010-03-17 2011-03-16 Method for production of polymer containing fibres
CA2789647A CA2789647C (fr) 2010-03-17 2011-03-16 Procede de fabrication de fibres contenant un polymere
RU2012144053/05A RU2545331C2 (ru) 2010-03-17 2011-03-16 Способ получения полимерсодержащих волокон
US13/634,484 US20130172478A1 (en) 2010-03-17 2011-03-16 Method for production of polymer containing fibres
EP11709018.3A EP2547810B1 (fr) 2010-03-17 2011-03-16 Procédé de fabrication de fibres contenant un polymère

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
PCT/EP2010/001694 WO2011113446A1 (fr) 2010-03-17 2010-03-17 Procédé de fabrication de fibres contenant un polypeptide
EPPCT/EP2010/001694 2010-03-17

Publications (1)

Publication Number Publication Date
WO2011113592A1 true WO2011113592A1 (fr) 2011-09-22

Family

ID=43035008

Family Applications (2)

Application Number Title Priority Date Filing Date
PCT/EP2010/001694 WO2011113446A1 (fr) 2010-03-17 2010-03-17 Procédé de fabrication de fibres contenant un polypeptide
PCT/EP2011/001307 WO2011113592A1 (fr) 2010-03-17 2011-03-16 Procédé de fabrication de fibres contenant un polymère

Family Applications Before (1)

Application Number Title Priority Date Filing Date
PCT/EP2010/001694 WO2011113446A1 (fr) 2010-03-17 2010-03-17 Procédé de fabrication de fibres contenant un polypeptide

Country Status (5)

Country Link
US (1) US20130172478A1 (fr)
AU (1) AU2011229482B2 (fr)
CA (1) CA2789647C (fr)
RU (1) RU2545331C2 (fr)
WO (2) WO2011113446A1 (fr)

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014002605A1 (fr) 2012-06-28 2014-01-03 スパイバー株式会社 Fibre de protéine teintée dans la masse et procédé pour produire celle-ci
WO2016149414A1 (fr) 2015-03-16 2016-09-22 Bolt Threads, Inc. Fibres de soie améliorées
WO2016201369A1 (fr) 2015-06-11 2016-12-15 Bolt Threads, Inc. Fils de fibres de protéine recombinée ayant des propriétés améliorées
WO2017011679A1 (fr) 2015-07-14 2017-01-19 Silk Therapeutics, Inc. Pièce d'habillement technique en soie et procédés de préparation
WO2017025964A1 (fr) 2015-08-10 2017-02-16 Seevix Material Sciences Ltd. Compositions et procédés de fabrication de soie de fil de traîne d'araignée synthétique
US9617315B2 (en) 2011-06-01 2017-04-11 Spiber Inc. Artificial polypeptide fiber and method for producing the same
WO2017138002A1 (fr) 2016-02-11 2017-08-17 Seevix Material Sciences Ltd. Matériaux composites comprenant de la soie d'araignée synthétique de type fil de traîne
WO2017188430A1 (fr) * 2016-04-28 2017-11-02 Ｓｐｉｂｅｒ株式会社 Fibroïne modifiée
WO2017188434A1 (fr) * 2016-04-28 2017-11-02 Ｓｐｉｂｅｒ株式会社 Fibroïne modifiée
WO2017214618A1 (fr) 2016-06-10 2017-12-14 Bolt Threads, Inc. Fils de fibres de protéine recombinante ayant des propriétés améliorées
WO2018034111A1 (fr) * 2016-08-19 2018-02-22 国立研究開発法人理化学研究所 Composition de moulage composite comprenant une protéine de type fibroïne, et procédé de production de la composition de moulage composite
DE102016222480A1 (de) * 2016-11-16 2018-05-17 Adidas Ag Bekleidungsstück, das Spinnenseide aufweist oder Schuh, der Spinnenseide aufweist
JP6337252B1 (ja) * 2017-03-10 2018-06-06 Ｓｐｉｂｅｒ株式会社 高収縮人造フィブロイン繊維及びその製造方法、並びに人造フィブロイン繊維の収縮方法
WO2018164190A1 (fr) * 2017-03-10 2018-09-13 Ｓｐｉｂｅｒ株式会社 Fibres de fibroïne synthétique
WO2019022163A1 (fr) * 2017-07-26 2019-01-31 Ｓｐｉｂｅｒ株式会社 Fibroïne modifiée
WO2019151432A1 (fr) * 2018-01-31 2019-08-08 Ｓｐｉｂｅｒ株式会社 Procédé de préparation d'une fibre protéinique frisée d'adhérence à l'huile
WO2020033970A2 (fr) 2018-08-10 2020-02-13 Bolt Threads, Inc. Composition pour un corps moulé
CN112074631A (zh) * 2018-04-03 2020-12-11 长谷虎纺绩株式会社 混纺纱以及该混纺纱的织物和该织物的制造方法
CN112218979A (zh) * 2018-04-03 2021-01-12 丝芭博株式会社 复合纤维及其制造方法
US10975206B2 (en) 2015-04-09 2021-04-13 Spiber Inc. Polar solvent solution and production method thereof
US11174572B2 (en) 2015-08-20 2021-11-16 Riken Composite molding composition including fibroin-like protein, and method for producing composite molding composition
US11192982B2 (en) 2013-09-17 2021-12-07 Bolt Threads, Inc. Methods and compositions for synthesizing improved silk fibers
US11447532B2 (en) 2016-09-14 2022-09-20 Bolt Threads, Inc. Long uniform recombinant protein fibers
US11668024B2 (en) 2015-04-09 2023-06-06 Spiber, Inc. Polar solvent solution and production method thereof

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9907815B2 (en) * 2013-11-21 2018-03-06 The University Of Akron Method for preparation of filaments of poly(α-lipoic acid) polymers
JP6848173B2 (ja) * 2015-10-09 2021-03-24 東レ株式会社 含繊維結晶、含繊維結晶の製造方法、含繊維結晶の製造装置および薬剤ソーキング装置
CN113136635A (zh) * 2021-04-20 2021-07-20 徐建平 一种玉米肽纤维、制备方法及其应用
EP4252549A1 (fr) 2022-03-28 2023-10-04 Mirai Foods AG Procédés et compositions de préparation de faisceaux musculaires fibreux pour la production de viande d'élevage

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003057727A1 (fr) 2002-01-11 2003-07-17 Nexia Biotechnologies, Inc. Procedes de production de polypeptides de soie et de produits de ceux-ci
WO2003060099A2 (fr) * 2002-01-11 2003-07-24 Nexia Biotechnologies, Inc. Procedes et appareils de filage d'une proteine de soie d'araignee
WO2004000915A2 (fr) * 2002-06-24 2003-12-31 Tufts University Biomateriaux a base de soie et leurs methodes d'utilisation
EP1462548A1 (fr) * 2002-01-04 2004-09-29 Guanqi Li Fibre synthetique de phytoproteine et son procede de production
WO2006008163A2 (fr) 2004-07-22 2006-01-26 Technische Universitaet Muenchen Proteines de soies d'araignees recombinees
WO2006081749A1 (fr) * 2005-02-05 2006-08-10 Guanqi Li Solution de filage d'une fibre contenant des protéines, absorbant les ondes, faisant écran et absorbant la chaleur et procédé de préparation de celle-ci
WO2007025719A1 (fr) 2005-08-29 2007-03-08 Technische Universitaet Muenchen Protéines de soie arachnéenne modifiées
WO2008155304A1 (fr) 2007-06-20 2008-12-24 Basf Se Protéines synthétiques répétitives, leur fabrication et leur utilisation
US20090259010A1 (en) * 2006-12-12 2009-10-15 Hiking Group Co., Ltd. Modified polyacrylonitrile fiber and method of preparing the same

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5965125A (en) * 1995-10-25 1999-10-12 Transkaryotic Therapies, Inc. Hybrid matrix implants and explants
US8173772B2 (en) * 2005-12-30 2012-05-08 Spiber Technologies Ab Spider silk proteins and methods for producing spider silk proteins
US9427499B2 (en) * 2008-11-17 2016-08-30 Trustees Of Tufts College Surface modification of silk fibroin matrices with poly(ethylene glycol) useful as anti-adhesion barriers and anti-thrombotic materials

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1462548A1 (fr) * 2002-01-04 2004-09-29 Guanqi Li Fibre synthetique de phytoproteine et son procede de production
WO2003057727A1 (fr) 2002-01-11 2003-07-17 Nexia Biotechnologies, Inc. Procedes de production de polypeptides de soie et de produits de ceux-ci
WO2003060099A2 (fr) * 2002-01-11 2003-07-24 Nexia Biotechnologies, Inc. Procedes et appareils de filage d'une proteine de soie d'araignee
WO2004000915A2 (fr) * 2002-06-24 2003-12-31 Tufts University Biomateriaux a base de soie et leurs methodes d'utilisation
WO2006008163A2 (fr) 2004-07-22 2006-01-26 Technische Universitaet Muenchen Proteines de soies d'araignees recombinees
WO2006081749A1 (fr) * 2005-02-05 2006-08-10 Guanqi Li Solution de filage d'une fibre contenant des protéines, absorbant les ondes, faisant écran et absorbant la chaleur et procédé de préparation de celle-ci
WO2007025719A1 (fr) 2005-08-29 2007-03-08 Technische Universitaet Muenchen Protéines de soie arachnéenne modifiées
US20090259010A1 (en) * 2006-12-12 2009-10-15 Hiking Group Co., Ltd. Modified polyacrylonitrile fiber and method of preparing the same
WO2008155304A1 (fr) 2007-06-20 2008-12-24 Basf Se Protéines synthétiques répétitives, leur fabrication et leur utilisation

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
"Helvetica Chimica Acta", 1995, article "A multilingual glossary of biotechnological terms: (IUPAC Recommendations)"
ELVIN ET AL., NATURE, 2005, pages 999 - 1002
EXLER ET AL., ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2007, pages 3559 - 3562
GOSLINE ET AL., J. EXP. BIOL., 1999, pages 3295
HUEMMERICH ET AL., CURR. BIOL., 2004, pages 2070 - 4
RAMMENSEE ET AL., PNAS, 2008, pages 6590 - 6595
SCHEIBEL, MICROB. CELL. FACT., 2004, pages 14

Cited By (53)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9617315B2 (en) 2011-06-01 2017-04-11 Spiber Inc. Artificial polypeptide fiber and method for producing the same
WO2014002605A1 (fr) 2012-06-28 2014-01-03 スパイバー株式会社 Fibre de protéine teintée dans la masse et procédé pour produire celle-ci
US9689089B2 (en) 2012-06-28 2017-06-27 Spiber Inc. Solution-dyed protein fiber and method for producing same
EP2868782B1 (fr) 2012-06-28 2020-07-15 Spiber Inc. Fibre de protéine teintée dans la masse et procédé pour produire celle-ci
US11192982B2 (en) 2013-09-17 2021-12-07 Bolt Threads, Inc. Methods and compositions for synthesizing improved silk fibers
US11505654B2 (en) 2013-09-17 2022-11-22 Bolt Threads, Inc. Methods and compositions for synthesizing improved silk fibers
WO2016149414A1 (fr) 2015-03-16 2016-09-22 Bolt Threads, Inc. Fibres de soie améliorées
AU2016233266B2 (en) * 2015-03-16 2020-08-27 Bolt Threads, Inc. Improved silk fibers
US10975206B2 (en) 2015-04-09 2021-04-13 Spiber Inc. Polar solvent solution and production method thereof
US11668024B2 (en) 2015-04-09 2023-06-06 Spiber, Inc. Polar solvent solution and production method thereof
WO2016201369A1 (fr) 2015-06-11 2016-12-15 Bolt Threads, Inc. Fils de fibres de protéine recombinée ayant des propriétés améliorées
WO2017011679A1 (fr) 2015-07-14 2017-01-19 Silk Therapeutics, Inc. Pièce d'habillement technique en soie et procédés de préparation
AU2016305360B2 (en) * 2015-08-10 2021-11-04 Seevix Material Sciences Ltd. Compositions and methods for fabricating synthetic dragline spider silk
WO2017025964A1 (fr) 2015-08-10 2017-02-16 Seevix Material Sciences Ltd. Compositions et procédés de fabrication de soie de fil de traîne d'araignée synthétique
EP3334754A4 (fr) * 2015-08-10 2019-01-16 Seevix Material Sciences Ltd. Compositions et procédés de fabrication de soie de fil de traîne d'araignée synthétique
US11174572B2 (en) 2015-08-20 2021-11-16 Riken Composite molding composition including fibroin-like protein, and method for producing composite molding composition
JP2019510541A (ja) * 2016-02-11 2019-04-18 シービックス マテリアル サイエンシーズ リミテッド 合成のクモドラグラインシルクを含む複合材料
US12030919B2 (en) 2016-02-11 2024-07-09 Seevix Material Sciences Ltd. Composite materials comprising synthetic dragline spider silk
JP2023017893A (ja) * 2016-02-11 2023-02-07 シービックス マテリアル サイエンシーズ リミテッド 合成のクモドラグラインシルクを含む複合材料
WO2017138002A1 (fr) 2016-02-11 2017-08-17 Seevix Material Sciences Ltd. Matériaux composites comprenant de la soie d'araignée synthétique de type fil de traîne
US11345727B2 (en) 2016-02-11 2022-05-31 Seevix Material Sciences Ltd. Composite materials comprising synthetic dragline spider silk
JP7176736B2 (ja) 2016-02-11 2022-11-22 シービックス マテリアル サイエンシーズ リミテッド 合成のクモドラグラインシルクを含む複合材料
JP7529300B2 (ja) 2016-02-11 2024-08-06 シービックス マテリアル サイエンシーズ リミテッド 合成のクモドラグラインシルクを含む複合材料
CN109477252A (zh) * 2016-02-11 2019-03-15 赛威克斯材料科学公司 包括合成拖丝蜘蛛丝的复合材料
US20190135880A1 (en) * 2016-04-28 2019-05-09 Spiber Inc. Modified Fibroin
JPWO2017188434A1 (ja) * 2016-04-28 2018-07-26 Ｓｐｉｂｅｒ株式会社 改変フィブロイン
CN109311954A (zh) * 2016-04-28 2019-02-05 丝芭博株式会社 改造丝心蛋白
JPWO2017188430A1 (ja) * 2016-04-28 2018-08-30 Ｓｐｉｂｅｒ株式会社 改変フィブロイン
WO2017188434A1 (fr) * 2016-04-28 2017-11-02 Ｓｐｉｂｅｒ株式会社 Fibroïne modifiée
WO2017188430A1 (fr) * 2016-04-28 2017-11-02 Ｓｐｉｂｅｒ株式会社 Fibroïne modifiée
CN109071619A (zh) * 2016-04-28 2018-12-21 丝芭博株式会社 改造丝心蛋白
WO2017214618A1 (fr) 2016-06-10 2017-12-14 Bolt Threads, Inc. Fils de fibres de protéine recombinante ayant des propriétés améliorées
WO2018034111A1 (fr) * 2016-08-19 2018-02-22 国立研究開発法人理化学研究所 Composition de moulage composite comprenant une protéine de type fibroïne, et procédé de production de la composition de moulage composite
US11447532B2 (en) 2016-09-14 2022-09-20 Bolt Threads, Inc. Long uniform recombinant protein fibers
DE102016222480A1 (de) * 2016-11-16 2018-05-17 Adidas Ag Bekleidungsstück, das Spinnenseide aufweist oder Schuh, der Spinnenseide aufweist
DE102016222480B4 (de) * 2016-11-16 2020-02-13 Adidas Ag Bekleidungsstück, das Spinnenseide aufweist oder Schuh, der Spinnenseide aufweist, und ein entsprechendes Herstellungsverfahren
JP2018150637A (ja) * 2017-03-10 2018-09-27 Ｓｐｉｂｅｒ株式会社 高収縮人造フィブロイン繊維及びその製造方法、並びに人造フィブロイン繊維の収縮方法
WO2018164190A1 (fr) * 2017-03-10 2018-09-13 Ｓｐｉｂｅｒ株式会社 Fibres de fibroïne synthétique
CN110475917A (zh) * 2017-03-10 2019-11-19 丝芭博株式会社 高收缩人造丝心蛋白纤维及其制造方法、以及人造丝心蛋白纤维的收缩方法
JP6337252B1 (ja) * 2017-03-10 2018-06-06 Ｓｐｉｂｅｒ株式会社 高収縮人造フィブロイン繊維及びその製造方法、並びに人造フィブロイン繊維の収縮方法
WO2018164021A1 (fr) * 2017-03-10 2018-09-13 Ｓｐｉｂｅｒ株式会社 Fibres de fibroïne synthétique à retrait élevé ainsi que procédé de fabrication de celles-ci, et procédé de rétraction de fibres de fibroïne synthétique
US12018056B2 (en) 2017-03-10 2024-06-25 Spiber Inc. Highly contracted synthetic fibroin fiber, production method therefor, and method for contracting synthetic fibroin fiber
US11692015B2 (en) 2017-07-26 2023-07-04 Spiber Inc. Modified fibroin
WO2019022163A1 (fr) * 2017-07-26 2019-01-31 Ｓｐｉｂｅｒ株式会社 Fibroïne modifiée
CN111051334A (zh) * 2017-07-26 2020-04-21 丝芭博株式会社 改造丝心蛋白
JPWO2019022163A1 (ja) * 2017-07-26 2020-02-06 Ｓｐｉｂｅｒ株式会社 改変フィブロイン
JP2020150945A (ja) * 2017-07-26 2020-09-24 Ｓｐｉｂｅｒ株式会社 改変フィブロイン
JP7048058B2 (ja) 2017-07-26 2022-04-05 Ｓｐｉｂｅｒ株式会社 改変フィブロイン
JP2020023580A (ja) * 2017-07-26 2020-02-13 Ｓｐｉｂｅｒ株式会社 改変フィブロイン
WO2019151432A1 (fr) * 2018-01-31 2019-08-08 Ｓｐｉｂｅｒ株式会社 Procédé de préparation d'une fibre protéinique frisée d'adhérence à l'huile
CN112218979A (zh) * 2018-04-03 2021-01-12 丝芭博株式会社 复合纤维及其制造方法
CN112074631A (zh) * 2018-04-03 2020-12-11 长谷虎纺绩株式会社 混纺纱以及该混纺纱的织物和该织物的制造方法
WO2020033970A2 (fr) 2018-08-10 2020-02-13 Bolt Threads, Inc. Composition pour un corps moulé

Also Published As

Publication number Publication date
AU2011229482A1 (en) 2012-08-30
RU2545331C2 (ru) 2015-03-27
WO2011113446A1 (fr) 2011-09-22
RU2012144053A (ru) 2014-04-27
CA2789647C (fr) 2016-04-26
CA2789647A1 (fr) 2011-09-22
US20130172478A1 (en) 2013-07-04
AU2011229482B2 (en) 2015-05-07

Similar Documents

Publication Publication Date Title
AU2011229482B2 (en) Method for production of polymer containing fibres
Rammensee et al. Rheological characterization of hydrogels formed by recombinantly produced spider silk
US11248120B2 (en) Methods for producing high toughness silk fibres
JP5128943B2 (ja) 組換えスパイダーシルクタンパク質
US9051453B2 (en) Polypeptide solution, artificial polypeptide fiber production method and polypeptide purification method using same
EP2899204B1 (fr) Support pour culture cellulaire contenant des protéines de soie d'araignée et procédés de production de celles-ci
JP5584932B2 (ja) タンパク質繊維の製造方法
CN104884463B (zh) 亲水性重组蛋白的提取方法
US20190248847A1 (en) Engineered Spider Silk Proteins And Uses Thereof
EP2547810B1 (fr) Procédé de fabrication de fibres contenant un polymère
Debabov et al. Recombinant spidroins as the basis for new materials
US11597750B2 (en) Method for producing a condensed adhesive phase of silk fusion proteins
AU2018351873A1 (en) Single alpha chain collagens
Singh et al. Spinning and applications of spider silk
US9034816B2 (en) Biopolymer having excellent tensile strength, extensibility and toughness
Breslauer et al. 9.04-Silks
JP7177453B2 (ja) ポリペプチド、核酸、成形体、組成物及びその製造方法、並びに物性向上剤
Adachi Chemical functionalization and characterization of silk fibroin materials
Niemelä Essays on the Implications of an Employee-empowering Agile Management Approach on Management Control Elements
Välisalmi Exploring silk protein assembly mechanisms for high-performance materials
Vollrath et al. The route to synthetic silks
Machado et al. Construction and bioproduction of a “green” synthetic protein-based polymer exhibiting a smart behaviour

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11709018

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2011709018

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2011229482

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 6900/DELNP/2012

Country of ref document: IN

ENP Entry into the national phase

Ref document number: 2789647

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2011229482

Country of ref document: AU

Date of ref document: 20110316

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 13634484

Country of ref document: US

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2012144053

Country of ref document: RU