CN115057940A - Fusion protein, biological fiber containing fusion protein, preparation method and application thereof - Google Patents
Fusion protein, biological fiber containing fusion protein, preparation method and application thereof Download PDFInfo
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- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/10—Materials or treatment for tissue regeneration for reconstruction of tendons or ligaments
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- C07K2319/00—Fusion polypeptide
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Abstract
The invention belongs to the technical field of biological materials, and particularly relates to a fusion protein, a biological fiber containing the fusion protein, and a preparation method and application of the biological fiber. The high-strength high-toughness biological fiber comprises a glutaraldehyde covalent cross-linked fusion protein fiber prepared by wet spinning and a salmon sperm DNA/protein electrostatic assembly composite fiber collected manually. The Resilin-ELP fusion protein solution is extruded into a glutaraldehyde coagulation bath or mixed with salmon sperm DNA according to different charge ratios to prepare the fusion protein solution respectively. The covalent cross-linked fiber is composed of an imine bond network formed by aldehyde and amino through Schiff base reaction, the electrostatic assembly composite fiber is composed of a DNA framework and a protein flexible chain which are interacted through supermolecules, and the two protein fibers have excellent mechanical properties and good biocompatibility. The high-performance protein fiber prepared by the invention can be used in the biomedical field needing mechanical support materials, such as rat abdominal hernia repair and the like.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to a fusion protein, a biological fiber containing the fusion protein, and a preparation method and application of the biological fiber.
Background
Compared with chemical synthetic fibers, natural protein fibers (silk, spider silk, auricularia auricula and the like) have the advantages of no toxicity, biodegradability, support of cell growth and differentiation, high mechanical strength, strong bionic property and the like, and attract the attention of students. Based on the principle of bionics, the recombinant DNA technology is utilized to do a lot of work in the aspect of constructing bionic structural protein, in particular to the aspect of manufacturing advanced biomaterials with good structure and performance, including preparation of biological fibers, biological films, hydrogel, biological adhesives, even soft robots and the like. However, the mechanical properties of most recombinant biological fibers can hardly combine the characteristics of high strength and high toughness of natural protein fibers, which limits the application research of the recombinant biological fibers in various fields to a great extent. With the deepened understanding of the relationship between protein sequence-structure-function, we can not only artificially synthesize natural mechanical structure protein polymers, but also fuse various high-performance protein sequences to produce novel protein polymers with multiple advantages and high strength and high toughness mechanical properties.
In addition to mechanical structural proteins, the Resilin proteins composed of amorphous structures have attracted much attention due to their outstanding mechanical properties such as low rigidity, high ductility, excellent resilience, fatigue resistance, etc., which has led to the preparation of Resilin proteins into biomechanical protein hydrogels for use in tissue engineering and functional nanostructured materials. However, due to its low strength, high performance biofibers based on Resilin proteins with balanced strength and toughness have been rarely reported. On the one hand, the Resilin proteins have an inherent disordered structure, and lack the rigid crystal structure (β -sheets or α -helix) required for high strength, resulting in weaker tensile strength of the Resilin-based biomaterials. Furthermore, although the Resilin protein can effect solution-gel transition by photo-controlled dityrosine cross-linking, this process lacks critical timeliness. It is difficult to achieve conversion from protein solution to fiber in a short time by crosslinking. Therefore, the preparation of high-strength and high-toughness bionic fibers by utilizing Resilin protein has great challenge.
Disclosure of Invention
In view of this, the invention provides a fusion protein, a biological fiber containing the fusion protein, and a preparation method and application thereof. The biological fiber prepared by the fusion protein has excellent mechanical property and good biocompatibility.
In order to achieve the above object, the present invention provides the following technical solutions:
a fusion protein comprising at least one resin block and at least one ELP block;
the resin block comprises at least one of (1) to (3):
(1) a protein fragment with an amino acid sequence shown as SEQ ID No. 1;
(2) a protein which is obtained by substituting, deleting or adding one or more amino acids in the fragment shown in (1) and has the same or similar activity with the protein shown in (1);
(3) a protein having at least 80% homology with the amino acid sequence shown in (1) or (2);
the ELP block comprises at least one of (I) to (III):
(I) a protein fragment with an amino acid sequence shown as SEQ ID NO. 2;
(II) a protein which is obtained by substituting, deleting or adding one or more amino acids in the fragment shown in (I) and has the same or similar activity with the protein shown in (1);
(III) a protein having at least 80% homology with the amino acid sequence shown in (I) or (II).
In some embodiments, the amino acid sequence of the Resilin block is shown in SEQ ID NO. 1: VGSGGRPSDSYGAPGGGNP, respectively; the amino acid sequence of the ELP block is shown as SEQ ID NO. 2: VPGKG.
In the present invention, the Resilin block and the ELP block in the fusion protein can be linked in any number and order. In some preferred embodiments, the fusion protein comprises repeating units based on m units of resin block linked to n ELP blocksThat is, the fusion protein consists of the sequence (VGSGGRPSDSYGAPGGGNP) m (VPGKG) n The basic repeating unit of (a); wherein m is an integer of 1 to 40, and n is an integer of 1 to 200. In some embodiments, m is equal to 1, n is equal to 5, and the sequence of the base repeat unit is VGSGGRPSDSYGAPGGGNP(VPGKG) 5 。
In the invention, the number of the basic repeating units is 4-40. In some embodiments, the number of basic repeat units can be specifically 4, 8, 10, 12, 15, 18, 20, 24, 25, 28, 30, 33, 35, 37, or 40.
In some embodiments, the fusion protein of the invention is RE-24, and the amino acid sequence thereof is shown in SEQ ID NO. 3. In the invention, the C end of the fusion protein is also connected with a protein tag, and the protein tag is preferably a 6 XHis tag; other amino acids, such as linker peptides, may also be introduced between or at both ends of the resin block and the ELP block.
The fusion protein is obtained by constructing an expression vector containing a target gene by a genetic engineering method, then transforming the expression vector into engineering escherichia coli for expression and purification, and the related method can be carried out according to a conventional method in the field.
The invention also provides application of the fusion protein in preparation of biological fibers.
The invention also provides a biological fiber which is prepared by wet spinning the fusion protein and the protein cross-linking agent.
In the invention, the protein cross-linking agent is dialdehyde OCH (CH) 2 ) n CHO, including at least one of glutaraldehyde, PEG dialdehyde, glyoxal. In some embodiments, the protein crosslinking agent is glutaraldehyde.
The invention also provides a preparation method of the biological fiber, which comprises the following steps:
A) dissolving the fusion protein in ultrapure water to obtain a protein solution;
B) mixing a cross-linking agent and water to obtain a coagulation bath spinning solution;
C) and adding the protein solution into a coagulating bath spinning solution for wet spinning, and collecting protein fibers.
The biological fiber provided by the invention is prepared by wet spinning the fusion protein and the protein cross-linking agent. The biological fiber is composed of an imine bond network formed by reacting aldehyde and amino through Schiff base.
In the preparation method, the concentration of the protein solution is 100-300 mg/mL. In some preferred embodiments, the concentration of the protein solution is 200-250 mg/mL; in some embodiments, the concentration of the protein solution may specifically be 200mg/mL, 210mg/mL, 220mg/mL, 230mg/mL, 240mg/mL, or 250 mg/mL.
The mass concentration of the coagulating bath spinning solution is 0.5-3 wt%, preferably 1-2 wt%; the pH value of the glutaraldehyde solution is controlled to be 6.5-8.0, and the preferable pH value is 7.0-7.4.
The protein solution was added using a syringe. Wherein the inner diameter of the needle head of the injector is 100-400 mu m, and the preferable range is 150-300 mu m; the adding speed of the protein solution is 5-50 mu L/min, and the preferable adding speed is 15-30 mu L/min.
The retention time of the protein solution fluid in the coagulating bath is 5-20 s, and the preferable retention time is 7-10; the collection speed is 2 to 5m/min, and more preferably 3 to 4 m/min.
The biological fiber prepared by the method is soaked in water and then subjected to post-stretching treatment. The post-stretching treatment degree of the protein fiber is 0-120% of the length of the protofilament; more preferably 50 to 100% of the length of the strand.
The invention also provides another biological fiber prepared from the fusion protein and salmon sperm DNA.
The invention also provides a preparation method of the biological fiber, which comprises the following steps:
A) dissolving the fusion protein in ultrapure water to obtain a protein solution;
B) dissolving salmon sperm DNA in ultrapure water to obtain a salmon sperm DNA solution;
C) and mixing the protein solution and the salmon sperm DNA solution, shaking, uniformly mixing, centrifuging to obtain a protein-DNA aggregate, and picking out the aggregate at a constant speed by using a syringe needle to obtain the composite fiber.
The preparation method provided by the invention comprises the following steps:
the concentration of the protein solution is 100-300 mg/mL, and preferably 100-150 mg/mL.
The concentration of the salmon sperm DNA solution is 5-20 mg/mL, and the preferable concentration is 10-15 mg/mL.
The molar ratio of lysine to phosphate radical in salmon sperm DNA in the protein solution is 1:1 or 2: 1.
In the present invention, salmon sperm DNA is a common commercially available product.
The method for picking out the aggregate in the present invention is not particularly limited, but the aggregate is preferably picked out using a syringe, and more preferably a standard commercially available 5mL syringe needle. In the present invention, the speed of picking out the aggregates is 30 to 60cm/min, and more preferably 40 to 50 cm/min.
The biological fiber provided by the invention is prepared by electrostatic assembly of fusion protein and salmon sperm DNA, and consists of a DNA framework and a protein flexible chain which are interacted by supermolecules.
The invention also provides application of the biological fiber in preparing artificial tendons, artificial ligaments, tissue repair materials, tissue defect materials or wound repair materials.
The invention also provides a tissue engineering material which is prepared from the two biological fibers and auxiliary agents acceptable in the medical field.
The biological fiber prepared from the fusion protein and the protein cross-linking agent is composed of a compact dynamic imine bond network, the network structure enables the cross-linked fiber to have the characteristics of high strength, high toughness, stable mechanical property and the like, and the protein fiber shows good biocompatibility and excellent rebound resilience, can be used for abdominal hernia repair requiring mechanical support materials, has a remarkable application effect, and has a wide application prospect. In the biological fiber prepared by electrostatic assembly of the fusion protein and salmon sperm DNA, the DNA chain plays a role of a rigid skeleton, the amino group of the fusion protein chain is anchored on the phosphate radical through the electrostatic action, and the fusion protein chain plays a role of a flexible bridge for connecting the DNA, so that the composite fiber has excellent mechanical properties. The biological fiber prepared by the fusion protein has excellent mechanical property and good biocompatibility, and can be used as a high-performance biological new material such as artificial tendon, artificial ligament, tissue repair and the like in the field of biomedicine.
Drawings
FIG. 1 is a schematic diagram of a process for preparing RE-24 covalently crosslinked protein fibers of example 1;
FIG. 2 RE-24 collected from example 1 covalently cross-linked protein fibers;
FIG. 3 scanning electron microscope image of RE-24 covalently cross-linked protein fiber in example 1;
FIG. 4 is the fiber mechanical tensile curve of RE-24 covalently cross-linked protein in example 1;
FIG. 5 is a schematic diagram of a method for preparing RE-24/DNA electrostatic assembly composite fibers in example 2;
FIG. 6 RE-24/DNA collected in example 2 electrostatically assembled composite fibers;
FIG. 7 scanning electron microscope image of RE-24/DNA electrostatic assembly composite fiber in example 2;
FIG. 8 the RE-24/DNA (1:1) electrostatic assembly complex fiber mechanical tensile curve in example 2;
FIG. 9 the RE-24/DNA (2:1) electrostatic assembly complex fiber mechanical tensile curve in example 2;
FIG. 10 is a diagram of a web fabric made from RE-24 covalently cross-linked protein fibers of example 3;
fig. 11 is a graph of RE-24 mesh fabric used in rat abdominal hernia repair experiments in example 3.
Detailed Description
The invention provides a fusion protein, a biological fiber containing the fusion protein, and a preparation method and application thereof. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
In the present invention, English, Chinese and abbreviated letters of 20 amino acids are shown in Table 1:
TABLE 1
The reagents or apparatus used in the present invention are commercially available.
The invention is further illustrated by the following examples:
example 1 biological fibers prepared by chemical crosslinking of fusion proteins/crosslinkers
Step 1: selecting fusion protein as RE-24, wherein the amino acid sequence of the fusion protein is shown as SEQ ID NO.3 and shown in table 2, and purifying after expression of engineered escherichia coli to obtain freeze-dried RE-24 powder;
step 2: dissolving 250mg of freeze-dried RE-24 powder in 1mL of ultrapure water, and dissolving the powder with the concentration of 250mg/mL by shaking;
and 3, step 3: preparing a 1.0 wt% aqueous solution from a 50% purchased glutaraldehyde solution, and adjusting the pH of the solution to 7.0 by using sodium hydroxide;
and 4, step 4: sucking the protein solution by using a 3.0mL syringe, selecting a needle with the inner diameter of 200 mu m, extruding the protein solution into a beaker filled with 1.0 wt% glutaraldehyde aqueous solution at the speed of 15 mu L/min, controlling the time of the protein solution in a coagulating bath to be 5s, and then collecting the formed fibers at the speed of 3m/min, as shown in figure 1;
and 5: the collected dry fibers were soaked in water to be softened and taken out, and post-stretching treatment was performed by 100% using a machine.
TABLE 2
| Protein name | Molecular weight | Amino acid sequence |
| RE-24 | 100200 | MGQG[VGSGGRPSDSYGAPGGGNP(VPGKG) 5 VPG] 24 WH 6 |
Through the steps, the RE-24 covalent cross-linked protein fiber with high strength and high toughness can be obtained, and the collected fiber is shown in figure 2, and the cross-linked fiber is golden yellow; the scanned image of the fiber is shown in fig. 3, the fiber has smooth surface, uniform size and long-range ordered molecular structure; the tensile mechanical property of the fiber is shown in figure 4, the breaking strength of the fiber reaches about 550MPa, and the toughness reaches 130MJ m at most -3 。
EXAMPLE 2 composite biological fibers prepared by Electrostatic Assembly of protein/DNA
Step 1: selecting fusion protein as RE-24, as shown in SEQ ID NO.3, expressing by engineering Escherichia coli, and purifying to obtain lyophilized RE-24 powder;
and 2, step: dissolving 100mg of the freeze-dried RE-24 powder in 1mL of ultrapure water, and dissolving the powder with the concentration of 100mg/mL by shaking;
and step 3: dissolving 10mg of purchased salmon sperm DNA in 1mL of ultrapure water, and dissolving the salmon sperm DNA into the ultrapure water with the concentration of 10mg/mL by oscillation;
and 4, step 4: mixing the prepared protein solution and salmon sperm DNA solution according to the charge ratio of 1:1 or 2:1, shaking, mixing uniformly, centrifuging to obtain protein-DNA aggregate, and picking out the aggregate at constant speed by using a 5mL syringe needle to obtain composite fiber, wherein the preparation method is shown in figure 5;
through the steps, the RE-24/DNA electrostatic assembly composite fiber with high strength and toughness can be obtained, the collected fiber is shown in figure 6, the composite fiber is in a white cylindrical shape, the scanning image of the fiber is shown in figure 7, the fiber is uniform in size, about 10 mu m in diameter, smooth in surface appearance and free of defects. The tensile mechanical properties of the fibers are shown in fig. 8 and 9. When the charge ratio of protein to DNA is 1:1, the breaking strength of the composite fiber reaches about 210MPa, and the toughness reaches 160MJ m -3 (ii) a When the charge ratio of protein to DNA is 2:1, the breaking strength of the composite fiber reaches about 400MPa, and the toughness reaches 260MJ m -3 。
Example 3 high strength and toughness biological fibers based on fusion proteins
Step 1: selecting fusion protein as RE-24, wherein the amino acid sequence of the fusion protein is shown as SEQ ID NO.3, and purifying after expressing by engineering escherichia coli to obtain freeze-dried RE-24 powder;
step 2: dissolving 250mg of freeze-dried RE-24 powder in 1mL of ultrapure water, and dissolving the powder with the concentration of 250mg/mL by shaking;
and 3, step 3: preparing a 1.0 wt% aqueous solution from a 50% purchased glutaraldehyde solution, and adjusting the pH of the solution to 7.0 by using sodium hydroxide;
and 4, step 4: sucking up the protein solution by using a 3.0mL syringe, selecting a needle with the inner diameter of 200 μm, extruding the protein solution into a beaker filled with 1.0 wt% glutaraldehyde water solution at the speed of 15 μ L/min, controlling the time of the protein solution in a coagulation bath to be 5s, and then collecting the formed fibers at the speed of 3m/min, as shown in FIG. 1;
and 5: soaking the collected dry fibers in water, taking out after the fibers are softened, and performing 100% post-stretching treatment by using a machine;
step 6: twisting the post-stretched fibers into a group of 10 fibers to obtain fiber bundles for weaving the mesh fabric, and carrying out rat abdominal hernia repair experiments.
Through the steps, the golden yellow fiber net with the size of 1.2x1.2 cm is obtained, as shown in fig. 10, the prepared fiber net fabric is used for rat abdominal hernia repair experiments, as shown in fig. 11, through observation for 6 weeks, the fiber net fabric can successfully repair rat abdominal hernia without causing tissue adhesion, and the fiber is shown to have excellent biocompatibility and mechanical support performance.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Sequence listing
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Ala Pro Gly Gly Gly Asn Pro Val Pro Gly Lys Gly Val Pro Gly Lys
770 775 780
Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly
785 790 795 800
Val Pro Gly Val Gly Ser Gly Gly Arg Pro Ser Asp Ser Tyr Gly Ala
805 810 815
Pro Gly Gly Gly Asn Pro Val Pro Gly Lys Gly Val Pro Gly Lys Gly
820 825 830
Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val
835 840 845
Pro Gly Val Gly Ser Gly Gly Arg Pro Ser Asp Ser Tyr Gly Ala Pro
850 855 860
Gly Gly Gly Asn Pro Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val
865 870 875 880
Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro
885 890 895
Gly Val Gly Ser Gly Gly Arg Pro Ser Asp Ser Tyr Gly Ala Pro Gly
900 905 910
Gly Gly Asn Pro Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro
915 920 925
Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly
930 935 940
Val Gly Ser Gly Gly Arg Pro Ser Asp Ser Tyr Gly Ala Pro Gly Gly
945 950 955 960
Gly Asn Pro Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly
965 970 975
Lys Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Val
980 985 990
Gly Ser Gly Gly Arg Pro Ser Asp Ser Tyr Gly Ala Pro Gly Gly Gly
995 1000 1005
Asn Pro Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Lys
1010 1015 1020
Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Val Gly
1025 1030 1035 1040
Ser Gly Gly Arg Pro Ser Asp Ser Tyr Gly Ala Pro Gly Gly Gly Asn
1045 1050 1055
Pro Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly
1060 1065 1070
Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Val Gly Ser
1075 1080 1085
Gly Gly Arg Pro Ser Asp Ser Tyr Gly Ala Pro Gly Gly Gly Asn Pro
1090 1095 1100
Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Lys Gly Val
1105 1110 1115 1120
Pro Gly Lys Gly Val Pro Gly Lys Gly Val Pro Gly Trp His His His
1125 1130 1135
His His His
Claims (13)
1. A fusion protein comprising at least one resin block and at least one ELP block;
the Resilin block comprises at least one of (1) to (3):
(1) a protein fragment with an amino acid sequence shown as SEQ ID No. 1;
(2) a protein which is obtained by substituting, deleting or adding one or more amino acids in the fragment shown in (1) and has the same or similar activity with the protein shown in (1);
(3) a protein having at least 80% homology with the amino acid sequence shown in (1) or (2);
the ELP block comprises at least one of (I) to (III):
(I) a protein fragment with an amino acid sequence shown as SEQ ID NO. 2;
(II) a protein which is obtained by substituting, deleting or adding one or more amino acids in the fragment shown in (I) and has the same or similar activity with the protein shown in (1);
(III) a protein having at least 80% homology with the amino acid sequence shown in (I) or (II).
2. The fusion protein according to claim 1, characterized in that it is based on a repeating unit of m resin blocks linked to n ELP blocks; wherein m is an integer of 1 to 40, and n is an integer of 1 to 200.
3. The fusion protein of claim 2, wherein the number of the basic repeating units is 4 to 40; m is equal to 1 and n is equal to 5.
4. Use of the fusion protein of any one of claims 1 to 3 for the preparation of a biological fiber.
5. A bio-fiber prepared by wet spinning the fusion protein of any one of claims 1 to 3 and a protein crosslinking agent.
6. The biofiber of claim 5, wherein the protein crosslinking agent is at least one of glutaraldehyde, PEG dialdehyde, glyoxal.
7. The method for preparing a biofiber according to claim 5 or 6, comprising the steps of:
A) dissolving the fusion protein in ultrapure water to obtain a protein solution;
B) mixing a cross-linking agent and water to obtain a coagulation bath spinning solution;
C) and adding the protein solution into a coagulating bath spinning solution for wet spinning, and collecting protein fibers.
8. The method according to claim 7, wherein the concentration of the protein solution is 100 to 300 mg/mL; the mass concentration of the coagulating bath spinning solution is 0.5-3 wt%, and the pH value of the solution is controlled to be 6.5-8.0;
adding the protein solution by using a liquid propelling device, wherein the inner diameter of an extrusion opening of the liquid propelling device is 100-400 mu m, and the adding speed of the protein solution is 5-50 mu L/min;
the protein solution fluid is kept in the coagulating bath for 5-20 s, and the collection speed is 2-5 m/min.
9. A biofiber obtained from the fusion protein of any one of claims 1 to 3 and salmon sperm DNA.
10. The method of producing a biofiber of claim 9, comprising the steps of:
A) dissolving the fusion protein in ultrapure water to obtain a protein solution;
B) dissolving salmon sperm DNA in ultrapure water to obtain a salmon sperm DNA solution;
C) and mixing the protein solution and the salmon sperm DNA solution, shaking, uniformly mixing, centrifuging to obtain a protein-DNA aggregate, and picking out the aggregate at a constant speed by using a syringe needle to obtain the composite fiber.
11. The production method according to claim 10,
the concentration of the protein solution is 100-300 mg/mL;
the concentration of the salmon sperm DNA solution is 5-20 mg/mL;
the molar ratio of lysine in the protein solution to phosphate radicals in salmon sperm DNA is 1:1 or 2: 1;
the speed of picking out the aggregates is 30-60 cm/min.
12. Use of the recombinant protein according to any one of claims 1 to 3, the biofabric according to claim 5 or 6, the biofabric produced by the production method according to claim 7 or 8, the biofabric according to claim 9, or the biofabric produced by the production method according to claim 10 or 11 for producing an artificial tendon, an artificial ligament, a tissue repair material, a tissue defect material, or a wound repair material.
13. A tissue engineering material comprising the biofiber of claim 5 or 6, the biofiber produced by the production method of claim 7 or 8, the biofiber of claim 9 or the biofiber produced by the production method of claim 10 or 11, and a medically acceptable auxiliary.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117756925A (en) * | 2023-12-26 | 2024-03-26 | 广州普言生物科技有限公司 | Recombinant elastin Pro.ELP, and preparation method and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117756925A (en) * | 2023-12-26 | 2024-03-26 | 广州普言生物科技有限公司 | Recombinant elastin Pro.ELP, and preparation method and application thereof |
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