WO2012175153A2 - Solution ou dispersion protéique aqueuse stable, procédé de préparation de la solution ou dispersion aqueuse stable, procédé de production de structures planes ou façonnées, d'imprégnations ou de revêtements à base de la solution ou dispersion protéique aqueuse stable, et application correspondante - Google Patents

Solution ou dispersion protéique aqueuse stable, procédé de préparation de la solution ou dispersion aqueuse stable, procédé de production de structures planes ou façonnées, d'imprégnations ou de revêtements à base de la solution ou dispersion protéique aqueuse stable, et application correspondante Download PDF

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Publication number
WO2012175153A2
WO2012175153A2 PCT/EP2012/001706 EP2012001706W WO2012175153A2 WO 2012175153 A2 WO2012175153 A2 WO 2012175153A2 EP 2012001706 W EP2012001706 W EP 2012001706W WO 2012175153 A2 WO2012175153 A2 WO 2012175153A2
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WO
WIPO (PCT)
Prior art keywords
protein
solution
dispersion
aqueous
stable
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PCT/EP2012/001706
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German (de)
English (en)
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WO2012175153A3 (fr
Inventor
Wiebke SCHMITZ
Gunter Scharfenberger
Ulrike Herrlich
Artem Davidenko
Original Assignee
Carl Freudenberg Kg
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Application filed by Carl Freudenberg Kg filed Critical Carl Freudenberg Kg
Publication of WO2012175153A2 publication Critical patent/WO2012175153A2/fr
Publication of WO2012175153A3 publication Critical patent/WO2012175153A3/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/0208—Tissues; Wipes; Patches
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • 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
    • C07K14/43513—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae
    • C07K14/43518—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae from spiders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/10—General cosmetic use

Definitions

  • Stable, aqueous protein solution or protein dispersion process for the preparation of the stable, aqueous solution or dispersion and
  • the present invention relates to a stable, aqueous solution or a stable, aqueous dispersion with at least one self-assembling protein. Furthermore, the invention relates to a process for the preparation of the stable, aqueous protein solution or protein dispersion, a process for the electro- or rotor spinning of self-assembling protein using the stable, aqueous protein solution or protein dispersion, process for the preparation of mold - or fabrics, of
  • Impregnations or coatings of the stable, aqueous protein solution or protein dispersion and the use of the stable, aqueous protein solution or protein dispersion or of the moldings or fabrics, impregnations or coatings produced therefrom.
  • shaped or flat structures are here in particular fibers
  • CONFIRMATION COPY Under a fiber structure is here in particular an orderly or disorderly single or multi-layered assembly of a plurality of fibers understood or three-dimensional arrangements of the fibers or a combination with carriers.
  • Preferred fiber structures are nonwovens here.
  • the self-assembling proteins are composed of polypeptides. These are made up of amino acids, especially from the 20 of course
  • the amino acids can be modified, for example acetylated.
  • the self-assembling proteins are in particular intrinsically unfolded proteins or microbead-forming proteins.
  • Exemplary proteins are silk-like proteins, silk proteins or spider silk proteins.
  • the proteins may be of natural origin or synthetic, for example by recombinant production.
  • the self-assembling proteins have the property that they are unstable in concentrated, aqueous solutions or dispersions. Due to the hydrophobic parts, the protein precipitates or the solution gels. As a result, the proteins from aqueous solution or dispersion usually can not be processed into fibers.
  • JP 2010 270426 A a method is described in which
  • Silk proteins of concentrated formic acid, hexafluoroacetone hydrate or hexafluoroisopropanol Electrospinning takes place at elevated temperature to prevent gelling of the spinning solution. However, this may result in prolonged, continuous processing of damage to the protein.
  • organic solvents is from aspects of
  • JP 2010-150712 describes the electrospinning of an aqueous
  • Silkworm silk protein solution with the addition of a nonionic surfactant and a water-soluble polymer.
  • the addition of nonionic surfactant may be of concern to health in medical applications.
  • the object of the present invention has been found to provide or produce a health-safe and / or a particularly environmentally friendly and particularly stable and highly concentrated solution or dispersion of at least one self-assembling protein, the efficient or improved production of pure protein molds or Fabrics, protein impregnations or coatings with a wide range of applications allows.
  • the concentrated, aqueous protein solutions or protein dispersions should be able to be prepared without additional additives that stabilize the protein in aqueous solution or dispersion.
  • the addition of additives to stabilize the proteins in aqueous solution can be problematic for later use, for example, in the medical field, since the additives themselves must be toxicologically safe.
  • Rotor spinning process allows or improved. The same applies to the production of pure protein foams, gels, films, films or of pure protein impregnations or coatings.
  • Impregnations particularly suitable for the medical field preferably for or as wound dressings, surgical sutures or as a biocompatible coating of surfaces, in particular fibers,
  • Fibrous structures such as yarns or nonwovens, tissue engineering scaffolds, medical textiles, tubing, implants or foams, as hypoallergenic surfaces, for cosmetic products, for hygiene products, for household products and / or for filtration applications, for example tobacco and engine filters ,
  • the protein solution or protein dispersion in an aqueous solution or dispersion comprises at least one physically stabilized self-assembling protein having a protein concentration of from 5% by weight to 35% by weight, preferably from 5% by weight to 30% by weight. %, more preferably from 5 wt .-% to 25 wt .-% based on the total weight of the stable solution or dispersion.
  • the stable protein solution or protein dispersion according to the invention is understood to mean a stability of at least 5 hours, preferably more than 12 hours, more preferably more than 20 hours or even more than 48 hours.
  • a chemical stabilizer additive is currently not provided or required.
  • the water base is a particularly environmentally friendly solution.
  • water as a base for example for the
  • Spinning solution can be dispensed with the use of organic solvents. This represents from the perspective of work safety and the
  • pure, aqueous protein solutions or dispersions can be prepared in very high concentration and with high stability by the invention.
  • the self-assembling protein is physically stabilized in an aqueous solution or dispersion,
  • Room temperature ie in particular above 20 ° C, preferably between 30 ° C and 90 ° C, more preferably between 30 ° C and 50 ° C.
  • a stabilization sufficient for the processing of the protein solution or protein dispersion, ie without protein breakdown, is achieved already at about 30 ° C. It is furthermore surprising that it is preferably sufficient to carry out the dialysis for 3 to 4 hours at 30 ° C. and then at room temperature. This is a particularly protein-friendly process is realized.
  • Stabilizer additive pure, aqueous protein solutions or protein dispersions in very high concentration and with high stability can be produced. These can be processed by means of spinning processes into fibers or fiber structures, in particular nonwovens, in particular by means of electrical or
  • the self-assembling protein of the stable, aqueous protein solution or protein dispersion may be of natural origin or synthetic, in particular by recombinant production constructed from repetitive units of a structural protein, in particular an insect protein, preferably resilin, and / or a spider silk protein or the like or thereof derived protein, in particular with insertion or attachment of oligoamino acid blocks and / or an intrinsically unfolded protein or a microbead-forming protein.
  • the protein can thus also be a combination of the proteins derived from natural structural protein sequences, in particular a combination of the sequence of one protein with the sequence of another structural protein, particularly preferably a combination of the sequence of a spider silk with the sequence of resilin.
  • the self-assembling protein of the stable, aqueous protein solution or protein dispersion is preferably an R16, R16Arg 8 - (R16 protein with
  • the self-assembling proteins designated R16 and S16 and their preparation are described in detail, for example, in WO 2008/155304.
  • the protein and its production designated C16 are described in detail in WO 2007/082936.
  • additives or additives may optionally be added to the protein solution or protein dispersion.
  • the stable, aqueous protein solution or protein dispersion preferably comprises at least one additive selected from
  • Polymers for example in the form of dispersions, or preferably water-soluble polymers, such as polyalkylene oxides, such as polyethylene oxides, polyethylene glycols or polypropylene glycols, polyvinyl alcohols,
  • Polyethyleneimines polyvinylpyrrolidones, polyvinylamines, polyvinyl acetates, polyacrylates, polyalkylacrylates, such as polymethacrylates,
  • Polyhydroxyalkyl acrylates such as poly (2-hydroxymethacrylates) or poly (2-hydroxyethacrylates), polyvinylformamides, polyacrylamides,
  • Polycarboxylic acids such as polyacrylic acids or polymethacrylic acids, natural polymers such as galactomannan, chitosan, collagen, albumin, xanthan, alginates, gelatin, cellulose or starch, in particular their water-soluble derivatives such as hydroxypropyl cellulose, carboxymethyl starch, oxidized starch, carboxymethyl cellulose,
  • Carrier-forming polymers preferably combinations of two or more polymers, homo- and copolymers,
  • Processing aids such as spinning aids, preferably organic
  • Solvents such as alcohols, for example ethanol or butanol, and / or surface-active agents, such as surfactants for adjusting the surface tension or the vapor pressure of the solution or conductive salts for adjusting the conductivity,
  • Crosslinking agents for example as an addition to the spinning solution or for the subsequent crosslinking of fibers,
  • Anti-infective agents analgesics or antiviral additives, antiseptics, antiinflammatory agents, wound healing agents, haemostatic agents, proteins, protein fragments, peptides and / or peptide sequences, polysaccharides, for example chitosan,
  • Growth factors such as purines or pyrimidines, living cells, ß-tricalcium phosphate, hydroxyapatite, in particular specifically
  • Hydroxyapatite nanoparticles hydroxyapatite nanoparticles, antimicrobial agents, amino acids, enzymes, vitamins, antioxidants,
  • Skin or hair cosmetic agents for example effect pigments, and / or
  • odor adsorbing additives such as activated carbon or cyclodextrins
  • Crown ethers biologically active metals or metal alloys, protein adsorption-preventing substances, such as polyethylene oxide.
  • polymers can be used as homopolymers, as copolymers, for example as block copolymers, graft copolymers, comb copolymers or brush copolymers, star polymers, random or alternating
  • additives can be added or adsorbed in pure form, in any mixture with one another and / or in encapsulated form.
  • Polyethylene oxide polyethylene glycol, polyvinyl alcohol and / or
  • Polyvinylpyrrolidone which are dissolved as water-soluble polymers directly in the protein solution or protein dispersion and are toxicologically safe.
  • the additives are present in a concentration range from 0.01% by weight to 40% by weight, preferably in a concentration range from 0.01% by weight to 30% by weight and more preferably in one
  • the pharmacologically active substances or medicaments used as additives preferably also come in smaller amounts
  • Self-assembling protein preferably dissolved in a low concentration of about 0.5 wt.% To 4 wt .-% in a chaotropic salt, dialyzed or ultrafiltered with an agitation, in particular by stirring, against an aqueous buffer, gel-filtered or treated by other osmotic methods or the chaotropic salt is separated by precipitation, and the protein solution or Protein dispersion is concentrated with agitation, in particular by stirring, by heating to about 30 ° C to 90 ° C, preferably to about 50 ° C to 80 ° C.
  • the dialysis is carried out to remove the chaotropic salt from the protein solution or protein dispersion preferably against an aqueous buffer having a pH in the range of 4 to 14, more preferably in the pH range of 6 to 14 and most preferably in the pH Range from 7 to 12.
  • an aqueous buffer having a pH in the range of 4 to 14, more preferably in the pH range of 6 to 14 and most preferably in the pH Range from 7 to 12.
  • self-assembling protein preferably in a high concentration of about 7 wt .-% to 35 wt .-%, particularly preferably from 7 wt .-% to 30 wt .-%, most preferably from 7 wt .-% to 25 wt. %, dissolved in a chaotropic salt, with agitation, in particular by stirring, against a room temperature, in particular to about 30 ° C to 50 ° C, heated, aqueous buffer, which is optionally exchanged for distilled water, dialyzed or ultrafiltered, gel-filtered or treated by other osmotic methods, or the chaotropic salt is separated by precipitation.
  • the pH of the protein solution or protein dispersion of preferably alkaline buffer is lowered to a neutral level (about pH 6-7), which is particularly gentle for the Protein is.
  • Guanidinium thiocyanate, calcium chloride, lithium bromide, urea or an ionic liquid is preferably used as solubilizer as the chaotropic salt for the process for producing the stable, aqueous protein solution or protein dispersion according to the invention.
  • the chaoptropic salt for example guanidinium thiocyanate, is almost completely removed from the aqueous protein solution or protein dispersion. This gives a pure, aqueous solution of protein and water.
  • the salt for example guanidinium thiocyanate, could interfere with a subsequent spinning process, in particular electrospinning, because of its influence on the conductivity of the spinning solution, and could be toxic to the cells, especially when used in dressings.
  • the salt could also interfere with the formation and solidification of the fibers during the spinning process.
  • ultrafiltration As an alternative to dialysis of the protein solution or protein dispersion is also a gel filtration, ultrafiltration or other osmotic process into consideration. To concentrate the protein solution on a larger scale, ultrafiltration is particularly suitable.
  • the aqueous, highly concentrated protein solution or protein dispersion according to the invention has a very high stability, in particular a stability of at least 5 hours, preferably more than 12 hours, more preferably more than 20 hours or even more than 48 hours.
  • Protein solution can, without or on a shaped or sheet-like carrier, directly particularly pure protein-shaped or flat structures, ie protein fibers, fiber structures, in particular nonwovens, gels, films, films or foams, or particularly pure protein impregnations or protein coatings are produced.
  • the fibers or fiber structures, in particular nonwovens, are preferably produced by the spinning of the invention or
  • Stable, aqueous protein solution or protein dispersion prepared according to the invention without or on a shaped or flat-sheet carrier to the corresponding protein fibers or fiber structures, in particular - nonwovens.
  • shaped or sheet carriers are here or carrier
  • Fibers or fiber structures in particular nonwovens, understood or from gels, films, films or foams.
  • the spinning of the stable, aqueous solution with self-assembling protein is advantageously carried out by electric or rotor spinning.
  • the stabilization of the protein solution or protein dispersion prior to spinning can be carried out with a health-acceptable and / or a protein-sparing stabilization method.
  • Subsequent spinning is preferably possible at room temperature in a continuous process over an extended period of time without the protein solution or protein dispersion precipitating or gelling or being exposed to a harmful temperature effect.
  • the electro-spun fibers preferably have a fiber diameter of 1 nm to several micrometers, preferably from 10 nm to 2000 nm, more preferably a fiber diameter of 10 nm to 1000 nm and most preferably a fiber diameter of 10 nm to 800 nm.
  • the protein solution or dispersion according to the invention or prepared according to the invention can also be used as impregnation or coating, in particular by spray, dip or spincoating, without or on a sheet carrier can be used, for example of fibers or fiber structures, in particular nonwovens.
  • the stable, aqueous protein solution or dispersion according to the invention or prepared according to the invention can be used without or on a sheet carrier, preferably as a coating in gel, film or film form or in the form of patterns, such as dot or striped patterns, for example.
  • protein solution according to the invention or prepared according to the invention can be processed without or on a shaped or flat-sheet carrier into protein foams.
  • the protein solution or protein dispersion can thus be applied directly to a carrier, in particular to a shaped or flat structure, for example to a nonwoven fabric or another fiber structure, which or which
  • wound covering systems they can be spun onto a film, a film or a foam, for example when applied to a polyurethane foam in wound dressings.
  • a stable, pure, aqueous, highly concentrated protein solution or protein dispersion which is characterized by the high protein Concentration even without the use of spinning aids, such as viscosity boosters, spin.
  • additives for example those selected from agents for adjusting the viscosity, carrier-forming polymers, spinning auxiliaries, crosslinking agents,
  • Coating aids surface active agents, pharmacologically active agents, medicaments, wound healing agents, skin or hair cosmetic agents and / or odor adsorbing additives, before spinning or further processing. Furthermore, the stable, aqueous protein solution or dispersion according to the invention finds or find the fabrics produced therefrom.
  • Impregnations or coatings advantageously used for cosmetic products for example for creams, sponges, pads, strips or masks, for hygiene products, such as feminine hygiene, incontinence products, diapers, for household products, such as cleaning utensils, in particular
  • Cloths or sponges and / or for filter applications such as tobacco filters or engine filters.
  • a particularly preferred use is the stable, aqueous protein solution or protein dispersion according to the invention or find the molded or flat structures, impregnations or
  • Coatings for medical products preferably for or as
  • Wound dressings other textiles, tubes, fabric constructions or
  • the medical products can at least one of
  • Wound contact layer prepared according to a described method, which positively influences the wound healing process, or is designed as a biocompatible or hypoallergenic coating of surfaces. It is possible that the wound contact layer in contact with the wound may be absorbed by the body and broken down. In addition, it could prevent sticking to the wound. This will be painful
  • the abovementioned additives make fiber structures, in particular nonwovens, such as, for example, 3D-modeled nonwovens, gels, films, films or foams, or for the impregnations or coatings described in one of the above examples, for the shaped or flat structures, in particular the fibers Processes are produced, thus opening a particularly diverse scope.
  • nonwovens such as, for example, 3D-modeled nonwovens, gels, films, films or foams, or for the impregnations or coatings described in one of the above examples, for the shaped or flat structures, in particular the fibers Processes are produced, thus opening a particularly diverse scope.
  • the produced protein moldings or surfaces can be aftertreated,
  • R16Arg 8 protein (R16 protein with attached 8 amino acid oligoarginine block)
  • R16 protein (2 g) as a recombinant, self-assembling protein is dissolved in a chaotropic salt, in particular guanidinium thiocyanate solution (98 g 6M GuaSCN) with stirring.
  • a chaotropic salt in particular guanidinium thiocyanate solution (98 g 6M GuaSCN) with stirring.
  • the protein solution is transferred to a dialysis tube, and this is placed sealed in the buffer solution. During dialysis, the solution is stirred. The dialysis is preferably run for 8 to 14 hours, and the buffer is optionally replaced. Subsequently, the protein solution is taken out of the dialysis tube, placed in a tared beaker and weighed.
  • the protein solution is evaporated in an open beaker with stirring at about 70 ° C until a desired concentration of 7 wt .-% to 35 wt .-% R16 protein is reached.
  • the determination of the protein concentration is carried out by weighing.
  • the protein solution is processed into fibers by means of electrospinning at room temperature (high voltage 35 kV, electrode distance 10 cm).
  • the protein solution is prepared and dialyzed as in Example 1.
  • the protein solution is evaporated in an open beaker with stirring at 70 ° C until a desired concentration of 15 wt .-% to 20 wt .-% R16 protein is reached.
  • Protein concentration is by weighing.
  • the protein solution is processed into fibers by means of electrospinning at room temperature (high voltage 35 kV, electrode distance 10 cm).
  • R16Arge protein (450 mg) as a recombinant self-assembling protein is dissolved in a chaotropic salt, in particular guanidinium thiocyanate solution (3 ml of 6M GuaSCN, 15% m / v) with stirring.
  • a chaotropic salt in particular guanidinium thiocyanate solution (3 ml of 6M GuaSCN, 15% m / v) with stirring.
  • 1, 8 I sodium bicarbonate buffer are introduced and heated to about 50 ° C. The temperature is maintained during dialysis.
  • the protein solution is transferred to a dialysis tube, and this is placed sealed in the buffer solution. During dialysis, the solution is stirred.
  • the dialysis is preferably run for 8 to 14 hours, and the buffer is optionally replaced to a larger one
  • the concentration of the R16 protein is 7 wt .-% to 35 wt .-%.
  • the protein solution is processed to fibers with or without viscosity increasing agents by means of electrospinning at room temperature (high voltage 35 kV, electrode distance 10 cm).
  • the protein solution is prepared as in Example 3.
  • the protein solution is transferred to a dialysis tube, and this is placed sealed in the buffer solution. During dialysis, the solution is stirred. The dialysis is preferably allowed to proceed for 4 to 7 hours, the buffer is exchanged with 1.8 I of distilled water (pH 6 to 7), after which dialysis is continued for 4 to 7 hours. Subsequently, the protein solution is taken out of the dialysis tube.
  • the concentration of the R16 protein is 7 wt .-% to 35 wt .-%.
  • the protein solution is processed to fibers with or without viscosity increasing agents by means of electrospinning at room temperature (high voltage 35 kV, electrode distance 10 cm).
  • the protein solution is prepared as in Example 3 or 4, dialyzed and processed by electrospinning into fibers.
  • the protein solution is prepared as in Example 3, 4 or 5.
  • Example 5 Surprisingly, as in Example 5, a stabilization sufficient for processing the protein solution, ie without protein breakdown, is already achieved at 30 ° C. The temperature is maintained for 3 hours and then dialyzed further at room temperature.
  • the protein solution is added by electrospinning as in the previous examples with or without viscosity increasing agents
  • R16 protein (300 mg) as a recombinant self-assembling protein is dissolved in a chaotropic salt, in particular guanidinium thiocyanate solution (3 ml of 6M GuaSCN, 0% m / v) with stirring.
  • a chaotropic salt in particular guanidinium thiocyanate solution (3 ml of 6M GuaSCN, 0% m / v) with stirring.
  • guanidinium thiocyanate solution 3 ml of 6M GuaSCN, 0% m / v
  • the electro-spun R16 fibers of the examples preferably have a fiber diameter of 10 nm to 2000 nm, more preferably a fiber diameter of 10 nm to 1000 nm and most preferably a fiber diameter of 10 nm to 800 nm.
  • the preparation of the protein solution is carried out as in Example 1 with the addition of 4 wt .-% polyethylene oxide as an additive for adjusting the
  • the protein solution is prepared as in Example 2, dialyzed, concentrated and provided with 4% by weight of polyethylene oxide as a viscosity adjusting additive. Thereafter, the stable protein solution is processed by means of rotor spinning into fibers.
  • Example 9 The preparation of the protein solution is carried out as in Example 3, 5 or 6 with the addition of 15% by weight of polyvinylpyrrolidone as an additive for adjusting the viscosity. Thereafter, the protein solution is processed by means of rotor spinning into fibers
  • the preparation of the protein solution is carried out as in Example 1 with the optional addition of additives or viscosity-increasing agents.
  • the film or sheet is prepared from the protein solution by, for example, spin coating or doctoring.
  • the preparation of the protein solution is carried out as in Example 3, 5 or 6 with the optional addition of additives, in particular viscosity-increasing agents.
  • the protein solution is prepared and dialyzed as in Example 1.
  • the protein solution is evaporated in an open beaker with stirring at 70 ° C until a desired concentration of 5 wt .-% to 20 wt .-% R16 protein is reached.
  • the determination of the protein concentration is carried out by weighing.
  • the protein solution is uniformly sprayed on a viscose nonwoven fabric, for example by means of a spray gun with compressed air. Subsequently, the nonwoven fabric is dried at room temperature. Examples of the preparation of recombinant, self-assembling
  • the protein solution is prepared as in Example 12, dialyzed and
  • a viscose nonwoven fabric is dipped in the stable, concentrated protein solution and then dried at room temperature.
  • R16 protein or R16Arg 8 protein R16 protein with attached oligoarginine block.
  • the R16Arge protein is said to have special properties in wound healing.
  • the S16 protein or the C16 protein can be used.
  • the protein fibers, films, films or coatings are immersed for about 0.5 to 1 minute in ethanol or methanol and then dried at room temperature and to remove the solvent residues in a vacuum oven.
  • the protein fibers, films, films or coatings are immersed in potassium hydrogen phosphate buffer for about 2 hours and then washed with distilled water and dried at room temperature.

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Abstract

L'invention concerne la préparation d'une solution ou d'une dispersion fortement concentrée, non nocive pour la santé et/ou particulièrement neutre pour l'environnement et particulièrement stable, à base d'au moins une protéine apte à l'auto-assemblage, cette solution ou dispersion permettant une production efficace et améliorée de structures protéiques pures planes ou façonnées, d'imprégnations protéiques ou de revêtements protéiques, présentant un large champ d'application. Selon l'invention, la solution protéique ou la dispersion protéique dans une solution ou dispersion aqueuse comprend au moins une protéine apte à l'auto-assemblage physiquement stable présentant une concentration protéique de 5 à 35 % en poids, de préférence de 5 à 30 % en poids, mieux encore de 5 à 25 % en poids, relativement au poids total de la solution ou dispersion stable, présentant notamment une stabilité d'au moins 5 heures, de préférence de plus de 12 heures, mieux encore de plus de 20 heures et même de plus de 48 heures. Un additif de stabilisation chimique n'est ni prévu ni nécessaire.
PCT/EP2012/001706 2011-06-20 2012-04-20 Solution ou dispersion protéique aqueuse stable, procédé de préparation de la solution ou dispersion aqueuse stable, procédé de production de structures planes ou façonnées, d'imprégnations ou de revêtements à base de la solution ou dispersion protéique aqueuse stable, et application correspondante WO2012175153A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE201110105372 DE102011105372A1 (de) 2011-06-20 2011-06-20 Stabile, wässrige Protein-Lösung oder Protein-Dispersion, Verfahren zur Herstellung der stabilen, wässrigen Lösung oder Dispersion und Verfahren zur Herstellung von Form-oder Flächengebilden, Imprägnierungen oder Beschichtungen aus der stabilen, wässrigen Protein-Lösung oder Protein-Dispersion sowie deren Verwendung
DE102011105372.0 2011-06-20

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WO2012175153A2 true WO2012175153A2 (fr) 2012-12-27
WO2012175153A3 WO2012175153A3 (fr) 2013-05-30

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9757330B2 (en) 2013-10-18 2017-09-12 Industrial Technology Research Institute Recipe for in-situ gel, and implant, drug delivery system formed thereby
CN115466322A (zh) * 2022-09-26 2022-12-13 斐缦(长春)医药生物科技有限责任公司 一种自组装胶原蛋白及其制备方法和应用

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CN115466322B (zh) * 2022-09-26 2023-10-20 斐缦(长春)医药生物科技有限责任公司 一种自组装胶原蛋白及其制备方法和应用

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