WO2012175153A2

WO2012175153A2 - Stable aqueous protein solution or protein dispersion, method for the production of said stable aqueous solution or dispersion, and method for the production of molded articles or sheet material, impregnations, or coatings from the stable aqueous protein solution or protein dispersion, and use thereof

Info

Publication number: WO2012175153A2
Application number: PCT/EP2012/001706
Authority: WO
Inventors: Wiebke SCHMITZ; Gunter Scharfenberger; Ulrike Herrlich; Artem Davidenko
Original assignee: Carl Freudenberg Kg
Priority date: 2011-06-20
Filing date: 2012-04-20
Publication date: 2012-12-27
Also published as: WO2012175153A3; DE102011105372A1

Abstract

The aim of the invention is to prepare a particularly stable and highly concentrated solution or dispersion of at least one self-assembling protein, said solution or dispersion posing no health risk and/or being particularly environmentally friendly and allowing for efficient or improved production of molded articles or sheet material made of pure protein as well as protein impregnations or coatings in a large variety of fields of application. According to the invention, the protein solution or protein dispersion comprises, in an aqueous solution or dispersion, at least one physically stabilized self-assembling protein having a protein concentration of 5 to 35 wt.%, preferably 5 to 30 wt.%, especially preferably 5 to 25 wt.%, relative to the total weight of the stable solution or dispersion, in particular with a stability of at least 5 hours, preferably more than 12 hours, especially preferably more than 20 hours or even more than 48 hours. Note that no added stabilizer is included or required.

Description

Applicant: Carl Freudenberg KG, 69469 Weinheim

Stable, aqueous protein solution or protein dispersion, process for the preparation of the stable, aqueous solution or dispersion and

Process for the production of shaped or flat structures, impregnations or coatings from the stable, aqueous protein solution or protein dispersion and their use

Description 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.

Among shaped or flat structures are here in particular fibers,

Fiber structures, gels, films, films, foams or combinations thereof understood.

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

occurring amino acids, together. 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.

Prior Art From US Pat. No. 6,110,590, electrospinning is synthetic

produced silk from hexafluoroisopropanol known.

In 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. The use of organic solvents is from aspects of

Occupational safety and / or environmental protection also often not

desirable.

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, however, may be of concern to health in medical applications.

Presentation of the invention

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.

In addition, 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.

The concentrated protein solution or protein dispersion should also be characterized by a particularly high concentration and high stability

distinguished. In particular, the production of protein fibers or fiber structures, in particular nonwovens, by means of electric or

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.

Furthermore, the stable protein solution or dispersion or the moldings or fabrics, coatings or

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 ,

According to the invention, 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. By using 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

Environmentally friendly, especially with regard to the avoidance of harmful waste or by-products, a particular advantage.

Highly hydrophobic proteins have a very poor solubility in aqueous solution. Without stabilization, only dilute, aqueous protein solutions can be prepared. Due to the low concentration, these solutions or dispersions, for example, can not be processed into fibers without further additives, for example polymers. In addition, larger amounts of additives are needed in these dilute solutions.

On the other hand, pure, aqueous protein solutions or dispersions can be prepared in very high concentration and with high stability by the invention. For the process according to the invention for the preparation of the stable, aqueous protein solution or protein dispersion, the self-assembling protein is physically stabilized in an aqueous solution or dispersion,

in particular by mechanical movement and heating over

Room temperature, ie in particular above 20 ° C, preferably between 30 ° C and 90 ° C, more preferably between 30 ° C and 50 ° C. Surprisingly, 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.

Alone by the physical stabilization can without chemical

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

Rotor spinning. 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

attached oligoarginine block with 8 amino acids), S16 or C16 protein or the natural silk protein of the silkworm Bombyx mori or derived from the aforementioned proteins natural or synthetic protein.

The self-assembling proteins designated R16 and S16 and their preparation are described in detail, for example, in WO 2008/155304. For example, the protein and its production designated C16 are described in detail in WO 2007/082936.

Although physical stabilization does not require the addition of additives to stabilize the protein solution or protein dispersion.

Of course, however, 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

- Means for adjusting the viscosity, in particular water-insoluble

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,

- pharmacological agents or medicines, such as antibiotics,

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, 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.

The abovementioned 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

Systems, or be used in any mixture with each other.

The abovementioned additives can be added or adsorbed in pure form, in any mixture with one another and / or in encapsulated form.

Particularly preferred as a means for adjusting the viscosity

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. Furthermore, with regard to a use in the medical field as additives, preferably drugs and wound healing or hemostatic agents are used. Advantageously, 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

Concentration range of 2 wt .-% to 25 wt .-%. Depending on the application, the pharmacologically active substances or medicaments used as additives preferably also come in smaller amounts

Concentrations than those used above.

For the inventive method for producing the stable, aqueous protein solution or protein dispersion, the at least one

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. As an alternative to the aforementioned method, the at least one

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. By the optional replacement of the buffer solution by distilled water, the pH of the protein solution or protein dispersion of preferably alkaline buffer (greater than pH 7) 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. During dialysis, 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.

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.

From the stable, aqueous protein solution or protein dispersion according to the invention or the stable, aqueous

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.

Among the shaped or sheet carriers are here or carrier

Surfaces of 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. According to the method according to the invention, 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.

Furthermore, 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.

in particular by a pressure, doctor blade, roller or casting process, are used or processed.

A special application, for example, the coating

medical tubing with the protein solution or dispersion.

Furthermore, the 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

For example, in 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.

It is not necessary to use other substances, for example stabilizers for stabilizing the proteins in the protein solution. It lies

According to the invention, 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.

Regardless, can be used in the process for the production of moldings, impregnations or coatings from the

However, of course, the abovementioned 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

Tissue engineering scaffolds or implants. For example, the medical products can at least one

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

Dressing changes are avoided and the newly formed tissue layers in the wound are not destroyed by the change.

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.

It is based on the production of an environmentally friendly water-based and the increased concentration an effective method.

To stabilize against aqueous solutions, the produced protein moldings or surfaces can be aftertreated,

for example by immersion in ethanol or methanol or in

Potassium hydrogen phosphate buffer and washing with distilled water and subsequent drying.

Embodiment of the invention

The object of the invention will be explained in more detail by way of examples. The following materials are used:

- R 6 protein as a recombinant, self-assembling protein according to WO 2008/155304

R16Arg ₈ protein (R16 protein with attached 8 amino acid oligoarginine block)

Sodium bicarbonate buffer (10 mM, pH 10 to 12)

Guanidinium thiocyanate solution (6M); GuaSCN as a chaotropic salt

- Polyethylene oxide (MW 900000 g / mol) as an additive for adjusting the viscosity - poly-N-vinylpyrrolidone (MW 90000 g / mol) as an additive for adjusting the

viscosity

- dialysis tube (regenerated cellulose: MWCO (Molecular Weight Cut Off) from 10000-12000)

- Dialysis tubing clips

Examples of producing fibers from recombinant,

self-assembling proteins by electrospinning

example 1

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.

In a beaker 10 l sodium bicarbonate buffer are presented.

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).

Example 2

The protein solution is prepared and dialyzed as in Example 1.

Subsequently, 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. The determination of

Protein concentration is by weighing.

0.5% by weight to 3% by weight of polyethylene oxide (MW 900000 g / mol) as an additive for adjusting the viscosity are added and dissolved with stirring.

The protein solution is processed into fibers by means of electrospinning at room temperature (high voltage 35 kV, electrode distance 10 cm).

Example 3

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. In a beaker, 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

To achieve dilution factor. Subsequently, the protein solution is removed from the dialysis tube

removed. 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).

Example 4

The protein solution is prepared as in Example 3.

In a beaker, 1, 8 I sodium bicarbonate buffer are presented 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 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).

Example 5

The protein solution is prepared as in Example 3 or 4, dialyzed and processed by electrospinning into fibers.

Surprisingly, even at 30 ° C., sufficient stabilization for the processing of the protein solution, ie without protein breakdown, is achieved.

Example 6

The protein solution is prepared as in Example 3, 4 or 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.

Subsequently, the protein solution is added by electrospinning as in the previous examples with or without viscosity increasing agents

Room temperature further processed into fibers.

Example 7

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. Surprisingly, even at 30 ° C., sufficient stabilization for the processing of the protein solution, ie without protein breakdown, is achieved. Subsequently, the protein solution is dialyzed as in Example 5 and further processed by means of electrospinning at room temperature into fibers as in the abovementioned examples.

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.

Examples of producing fibers from recombinant,

self-assembling proteins by means of rotor spinning

Example 8

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

Viscosity.

Alternatively, 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

Examples of film or film production from recombinant,

Self-Assembling Proteins Example 10

The preparation of the protein solution is carried out as in Example 1 with the optional addition of additives or viscosity-increasing agents.

Thereafter, the film or sheet is prepared from the protein solution by, for example, spin coating or doctoring.

Example 11

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.

Thereafter, the film is removed from the protein solution, for example

Spin coating or by knife coating.

Examples of the preparation of nonwovens spray-coated with recombinant self-assembling proteins

Example 12

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

I

Proteins dip-coated nonwovens Example 13

The protein solution is prepared as in Example 12, dialyzed and

concentrated.

A viscose nonwoven fabric is dipped in the stable, concentrated protein solution and then dried at room temperature.

All experiments can be performed with R16 protein or R16Arg ₈ protein (R16 protein with attached oligoarginine block). The R16Arge protein is said to have special properties in wound healing.

Alternatively, the S16 protein or the C16 protein can be used.

Optional post-treatment of the manufactured protein fibers, protein films, protein films or protein coatings for stabilization against aqueous solutions 1st aftertreatment variant

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.

2nd aftertreatment variant

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.

Claims

claims

A stable, aqueous protein solution or protein dispersion, comprising in an aqueous solution or dispersion at least one physically stabilized self-assembling protein with a

Protein concentration of 5 wt .-% to 35 wt .-%, preferably from 5 wt .-% to 30 wt .-%, particularly preferably from 5 wt .-% to 25 wt .-% based on the total weight of the stable solution.

A stable, aqueous protein solution or protein dispersion according to claim 1, wherein the self-assembling protein is of natural origin or synthetic, in particular by recombinant production, constructed from repetitive units of a structural protein, in particular an insect protein and / or a spider silk protein or a similar or derived thereof Proteins, in particular with insertion or attachment of Oligoaminosäure blocks and / or an intrinsically unfolded protein or a microbead-forming protein.

A stable, aqueous protein solution or protein dispersion according to claim 1 or 2, wherein the self-assembling protein is an R16, R16Arge, S16 or C16 protein or a protein derived therefrom.

Stable, aqueous protein solution or protein dispersion according to one of the preceding claims, comprising at least one additive selected from viscosity adjusting agents, carrier-forming polymers, spinning aids, crosslinking agents, surface-active agents, pharmacologically active substances, medicaments,

Wound healing, skin or hair cosmetic

Active ingredients and / or odor adsorbing additives.

5. Stable, aqueous protein solution or protein dispersion according to claim 4, wherein the additive or the additives in a concentration of 0.01 Wt .-% to 40 wt .-% is present or present, preferably in one

Concentration of 0.01 wt .-% to 30 wt .-%, particularly preferably in a concentration of 2 wt .-% to 25 wt .-%.

6. A process for producing a stable, aqueous protein solution or protein dispersion according to any one of the preceding claims, wherein the at least one self-assembling protein is heated in an aqueous solution or dispersion above room temperature and mechanically agitated.

7. A process for the preparation of a stable, aqueous protein solution or protein dispersion according to claim 6, wherein the at least one self-assembling protein is dissolved in a concentration of 0.5 wt .-% to 4 wt .-% in a chaotropic salt, dialyzed with agitation against an aqueous buffer, ultrafiltered or 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 by heating to about 30 ° C to 90 ° C under motion.

8. A method for producing a stable, aqueous protein solution or protein dispersion according to claim 6, wherein the at least one self-assembling protein is dissolved in a concentration of 7 wt .-% to 35 wt .-% in a chaotropic salt, with agitation dialyzed against an aqueous buffer heated to about 30 ° C to 50 ° C, ultrafiltered or gel filtered or treated by other osmotic methods or the chaotropic salt is separated by precipitation.

9. A process for the preparation of a stable, aqueous protein solution or protein dispersion according to claim 7 or 8, wherein as a chaotropic salt Guanidinium thiocyanate, calcium chloride, lithium bromide, urea or an ionic liquid is used.

10. A process for the production of fibers or fiber structures, in particular of nonwovens, wherein a stable, aqueous protein solution or protein dispersion according to one of claims 1 to 5 used or produced by a method according to any one of claims 6 to 9, and then the protein solution or protein dispersion is spun into protein fibers or fiber structures, in particular nonwovens.

1 . Process for the electrospinning or rotor spinning of self-assembling protein, wherein a stable, aqueous solution or dispersion according to one of claims 1 to 5 is used or produced by a process according to any one of claims 6 to 9, and subsequently the protein solution or protein Dispersion is electrospun or rotorspun.

12. A process for the preparation of impregnations, feeds, gels, films or films, wherein a stable, aqueous protein solution or protein dispersion according to any one of claims 1 to 5 used or produced by a process according to any one of claims 6 to 9 , and then the protein solution or protein dispersion as a protein impregnation or protein coating, in particular by spray, dip or rotary coating of molding or

Sheet carriers is used or processed as a protein coating in gel, film or film form, in particular by a printing, doctor blade, roller or casting process, of shaped or sheet-like carriers.

13. Process for the production of molded or flat structures,

in particular fibers, fiber structures, gels, films, films or

Foaming, or of impregnations or coatings one of the preceding claims, wherein the protein solution or protein dispersion prior to spinning or processing at least one additive according to claim 4 or 5 is added. 14. Use of the stable, aqueous protein solution or protein dispersion or of the molded or flat structures produced therefrom, in particular fibers, fiber structures, gels, films, films or foams, or the impregnations or coatings produced therefrom according to any preceding claims for medical Products, preferred for or as wound dressings, tubes,

Tissue engineering or implants, for cosmetic products, for hygiene products, for household products and / or for filter applications. 15. Form or fabric, in particular fibers, fiber structures, gels, films, films or foams, or impregnations or

Coatings prepared by a process according to any one of claims 6 to 13.