55 results about "Spidroin" patented technology

The invention features methods and apparatuses for spinning silk protein fibers (biofilaments) from recombinant biofilament proteins. The methods are particularly useful for spinning fibers of spider silk or silkworm silk proteins from recombinant mammalian cells and may be used to spin such fibers for use in the manufacture of industrial and commercial products.

The invention features methods and apparatuses for spinning silk protein fibers (biofilaments) from recombinant biofilament proteins. The methods are particularly useful for spinning fibers of spider silk or silkworm silk proteins from recombinant mammalian cells and may be used to spin such fibers for use in the manufacture of industrial and commercial products.

The invention features methods and apparatuses for spinning silk protein fibers (biofilaments) from recombinant biofilament proteins. The methods are particularly useful for spinning fibers of spider silk or silkworm silk proteins from recombinant mammalian cells and may be used to spin such fibers for use in the manufacture of industrial and commercial products.

The invention discloses a method for preparing spider silk protein / polylactic acid composite nano-fiber yarn. The method comprises the following steps of preparing a spinning solution, preparing a bath solution and then adopting an improved electrostatic spinning method to guide, heat, wind and shape bunched fibers, so as to obtain the spider silk protein / polylactic acid composite nano-fiber yarn. The composite nano-fiber yarn of the invention can be continuously spun for more than 10 hours and have no broken ends, and a certain amount of spider silk protein effectively reduces the diameter of the fibers and improves the strength of the yarn. In addition, the fracture strength and flexibility of the composite nano-fiber yarn can be further improved through secondary stretching and combined twisting, so that the composite nano-fiber yarn can be applied in biomedical materials and other fields better.

The invention relates to natural polypeptides material for biomineralization and a preparation method thereof, in particular to a natural polypeptides material for preparing a crystal structure with fine biocompatibility by mineralization, belonging to the biological medical material field. The polypeptides material containing natural fibroin consists of degumming fibroin of insect silk and spidroin, which can be used for crystals mineralized at room temperature including zinc oxide, ferric oxide, gallium oxide, cobalt oxide, titanium oxide, hydroxylapatite, calcium phosphate and calcium carbonate. The silk fibroin solution after degumming is subjected to water washing, acid hydrolysis, dialysis, centrifuge enrichment to obtain the natural polypeptides material for mineralization, natural polypeptides material can be used for preparing composite materials with fine biocompatibility by mineralization.

A method of producing polymers of an isolated spider silk protein involves providing a solution of said spider silk protein in a liquid medium at pH 6.4 or higher and / or an ion composition that prevents polymerisation of the spider silk protein. The properties of the liquid medium are adjusted to a pH of 6.3 or lower and an ion composition that allows polymerisation of the spider silk protein. The spider silk protein is allowed to form polymers in the liquid medium, and the resulting spider silk protein polymers are isolated from the liquid medium. The resulting polymers are useful as fibers, films, foams, nets or meshes.

The invention relates to a recombinant spider silk protein preparation method.According to the method, full-length encoding genes of Araneus ventricosus minor ampullate spidroin(MiSp) and encoding genes of a full repeat region of pyriform spidroin(PySp) are taken as templates, gene segments are spliced to obtain a complete gene segment by a PCR fusion technique, the complete gene segment is connected with the pET28a carrier to construct recombinant plasmid, the recombinant plasmid is then transferred into BL21 for expression and is subjected to denaturation and purification to obtain recombinant spider silk protein solution, and the recombinant spider silk protein solution is subjected to freeze-drying preservation.The recombinant spider silk protein preparation method has the advantages of mild reaction condition, low cost, simple process and operational simplicity; the recombinant spider silk protein prepared by the method has good biocompatibility, high and predicable expression yield after being weighed, and is a potential protein fiber material having prospect of application based on industrialization.

The invention discloses recombinant spider silk protein, a coding gene thereof, an expression and purification method and application of the recombinant spider silk protein in the fields of silk formation, film formation and the like. The recombinant spider silk protein consists of an N-terminal non-repeated area (N-NRT), a core repeated area (REP), and a C-terminal non-repeated area (C-NRT), wherein the core repeated area consists of 1-3 ADF3Reps and / or 1-6 REPAs. A series of novel spider silk protein sequences designed by using the recombinant spider silk protein integrate the main characteristics, such as high strength and high elasticity, of spider silk. Moreover, the sequences are introduced into escherichia coli through a genetic engineering technology separately, a corresponding high-density fermentation process is established, the high-efficiency prokaryotic expression of the spider silk protein is achieved, and the fermentation yield can reach 0.5 g / L or above. In addition, aninventor develops a non-chromatographic protein purification method according to the properties of the spider silk protein, and the purity of the purified recombinant spider silk protein meets the requirements of silk formation or film formation.

A method of producing polymers of an isolated spider silk protein involves providing a solution of said spider silk protein in a liquid medium at pH 6.4 or higher and / or an ion composition that prevents polymerization of the spider silk protein. The properties of the liquid medium are adjusted to a pH of 6.3 or lower and an ion composition that allows polymerization of the spider silk protein. The spider silk protein is allowed to form polymers in the liquid medium, and the resulting spider silk protein polymers are isolated from the liquid medium. The resulting polymers are useful as fibers, films, foams, nets or meshes.

The invention provides a spidroin tissue repair material. The spidroin tissue repair material is membranoid substance containing spidroin, the inside average hole density is 1-2000 / mm<2>, and the average hole size is 1-300 [mu] m. The spidroin tissue repair material is characterized in that the membranoid substance contains the following raw materials in parts by weight: 80-90 parts of spidroin solution, 4-6 parts of spidroin nanofiber solution, 4-5 parts of paracetamol slow-release microcapsules, and 1-2 parts of essential oil. The invention relates to the fields of biological materials and spidroin. A biological material for repairing the damaged large tissue and complicated organs needs to have the capacity of promoting the formation of the blood vessels for enabling the large tissue and the complicated organs to have the biological function. But in the prior art, mostly, only the pure fibroin repair materials are available, the ideal blood vessel formation promoting effect and tissue repairing function are difficult to realize, while the tissue repair material prepared by adopting the scheme provided by the invention has the good tissue repairing effect.

The invention belongs to molecular biology and biomaterial fields, spider dragline silk gene is used as a base for artificial directed mutation of a part of amino acid sequence, a recombinant spider silk protein is obtained by an escherichia coli overexpression method and a Ni column purification technology, and after electrostatic spinning, a spider-silk-like protein is obtained, and the spider-silk-like protein has better consistency and elasticity than a natural spider silk protein. The spider-silk-like protein is an n-polyploid such as a tri-polyploid, hexa-polyploid or other integer-polyploid of an amino acid sequence shown in SEQ IDNO:5, wherein n is an integer equal to or greater than 1.

The invention relates to a mixed spidroin fiber preparing method.According to the method, the araneus ventricousus minor ampullate spidroin full-length coding gene and a piriform silk complete repetition area are taken as a template, splicing is conducted with the fusion PCR technique, connection with a pLX carrier is conducted, recombination quality construction is conducted, and expression is conducted in BL21.The degeneration purification means is adopted, and a mixed spidroin solution is obtained finally and stored after freeze-drying.Reaction conditions are mild, cost is low, the process is simple, and operation is easy.Prepared mixed spidroin fiber has high biocompatibility, uniform thickness and a smooth surface, and has potential application value in fields like biomedicine.

A protein structure capable of selective interaction with an organic target is provided. The protein structure is a polymer comprising as a repeating structural unit a recombinant fusion protein that is capable of selective interaction with the organic target. The fusion protein is comprising the moieties B, REP and CT, and optionally NT. B is a non-spidroin moiety of more than 30 amino acid residues, which provides the capacity of selective interaction with the organic target. REP is a moiety of from 70 to 300 amino acid residues and is derived from the repetitive fragment of a spider silk protein. CT is a moiety of from 70 to 120 amino acid residues and is derived from the C-terminal fragment of a spider silk protein. NT is an optional moiety of from 100 to 160 amino acid residues and is derived from the N-terminal fragment of a spider silk protein. The fusion protein and protein structure thereof is useful as an affinity medium and a cell scaffold material.

The invention discloses a preparation method and spinning of a high-concentration recombinant spider silk protein spinning solution. The molecular structure of a recombinant spider silk protein is NTD-(RP)n-CTD, an integer n represents the number of repetitions of an RP repetitive structural domain, and the amino acid sequence of the RP repetitive structural domain is shown as Seq ID No.1 or Seq ID No.2. The method can prepare the high-concentration recombinant spider silk protein spinning solution efficiently at low cost, and obtain artificial spider silks which are easy to modify and excellent in mechanical property.

The invention discloses MRRP protein, a gene, a carrier, engineering bacteria and application of the MRRP protein, the gene, the carrier and the engineering bacteria to preparation of wound healing medicine. The MRRP protein is formed by sequentially connecting mussel attachment protein Ma1, elastic drosophila melanogaster protein Res1, a spider silk protein nitrogen end sequence Nt1, a spider silk protein middle-piece repetitive sequence SSR, a spider silk protein carbon end sequence Ct1, elastic drosophila melanogaster protein Res2 and mussel attachment protein Ma2. The MRRP protein integrates self-assembling performance of spider silk protein, high elasticity of the elastic drosophila melanogaster protein and high viscosity of the mussel attachment protein, comprehensive performance is excellent, lengths are uniform, the MRRP protein can be biodegraded completely, the yield is high by adopting a microorganism method for preparing, and chemical pollution is avoided; the MRRP protein has high elasticity, and can bond wounds, so that the wounds are bonded without using lines.

The invention relates to the field of weaving of textiles and discloses a water-absorbing composite silk fabric and a processing method thereof. The method includes steps: subjecting modified polyethylene glycol to mixed reaction with fibroin; spinning and twisting to obtain modified silk fibers; mixing spider silk protein liquid with glucan; spinning and twisting to obtain modified spider silk fibers; blending the obtained modified silk fibers with the modified spider silk fibers to obtain a water-absorbing composite silk gray fabric; subjecting the water-absorbing composite silk gray fabricto mixed reaction with protease solution, performing high-temperature enzyme deactivation, adding chitosan mixed liquid, filtering after stirring reaction, and performing freeze drying to obtain the water-absorbing composite silk fabric. The water-absorbing composite silk fabric prepared according to the method is excellent in water absorbing performance and great in drapability.

A recombinant spider silk protein, consisting of no more than 800 amino acids, comprising a set of domains arranged according to the formula (NT)-REP-CT, wherein: the optional NT-domain, if present, comprises a sequence of 100 to 160 amino-acid residues derived from the N-terminal domain of a spider silk protein; the REP-domain comprises a sequence of 30 to 600 amino acid residues derived from therepetitive segment of a spider silk protein; and the CT-domain comprises a sequence of 70 to 120 amino acid residues derived from the C-terminal domain of a spider silk protein selected from: a sequence of 72 to 110 amino acid residues derived from the C-terminal domain of a spider silk protein, wherein the sequence comprises at least 7 residues independently selected from K, R, E and D; a sequence having at least 85% identity to SEQ ID NO: 15 or any one of SEQ ID NO:s 62-65 or 67-73; and a sequence having at least 70% identity to SEQ. ID NO: 64 or any one of SEQ ID NO:s 62-65 or 67-73, wherein the sequence comprises at least 7 residues independently selected from K, R, E and D. Solutions, polymers, uses and methods of manufacture thereof.

The invention discloses a ciprofloxacin dry powder inhaler and a preparation method thereof. The dry powder inhaler is prepared from a drug and a binary auxiliary material, wherein the drug accounts for 80 to 20 percent by mass of the dry powder inhaler, the binary auxiliary material accounts for 20 to 80 percent by mass of the dry powder inhaler, and the binary auxiliary material comprises a mucus diluting agent and biological adhesive protein; the mucus diluting agent is at least one of mannitol, ambroxol hydrochloride, aminothiopropionic acid, and bromhexine hydrochloride; and the biological adhesive protein is at least one of silk fibroin, albumin or spidroin. The dry powder inhaler prepared by the invention is good in mobility, good in aerodynamic performance, capable of targeting thedrug to lung and capable of effectively increasing the local concentration of the drug, has mucus diluting and biological mucosa adsorption characteristics, and is particularly suitable for a patientwith bronchiectasia caused by the chronic infection of the lung.

The invention relates to a nanoparticle emulsion loaded with a skin nutrition agent and a preparation method thereof. The nanoparticle emulsion is characterized by comprising the following ingredientsin percentage by weight: 0.1 percent to 50 percent of spider silk protein, 0.1 percent to 10 percent of bis[2-Hydroxyethyl] imino tris-(hydroxymethyl)-methane, 0.1 percent to 40 percent of the skin nutrition agent and 0.1 percent to 85 percent of a water and ethanol mixture, wherein the molecular weight of the spider silk protein is 80 to 100 kDa. The obtained nanoparticle emulsion loaded with the skin nutrition agent has higher biocompatibility, is milder, and has low irritation. Compared with other spider silk protein nanoemulsion, the nanoparticle emulsion has the advantages of higher encapsulation rate, better absorption permeability and higher stability.

The present invention discloses the drag silk performance of the cloud spider and its silk protein gene sequence. The silk protein amino acid sequence is shown in SEQ ID NO: 1 or / and SEQ ID NO: 2, or its amino acid sequence is shown in SEQ ID NO: 1 Or / and the amino acid sequence obtained by the polypeptide described in SEQ ID NO: 2 through substitution, deletion or addition of one or several amino acid residues and having equivalent mechanical properties. The present invention discovers the protein sequence and gene sequence of the drag silk proteins MaSp1 and MaSp2 of the cloud spider for the first time, and finds that the drag silk of the cloud spider has excellent mechanical properties and very strong toughness, which is significantly better than that of other spider species. The present invention lays the foundation for further understanding the evolution pattern of spidroin proteins of different species and different properties, and lays an important theory for the development of super spidroin proteins.

Transgenic silkworms stably expressing synthetic spider silk genes or composite silkworm / spider silk genes are disclosed. The exogenous spider silk genes are stably intergrated into a defined site of the fibroin heavy chain intron or a fibroin light chain intron of silkworms. Synthetic spider silk proteins and composite spider silk-silkworm genes and proteins are provided. The expression of exogenous spider silk genes is driven by the endogenous fibroin heavy chain promoter, improving the genetic stability of transgenic silkworms. The composite silkworm / spider silk fibers exhibit exceptional mechanical performance, compared to normal silkworm silk fibers and other transgenic silkworm fibers.

The invention provides an absorbable medical suture and a preparation method thereof in the technical field of medical materials. The raw material of the absorbable medical suture is modified spidroin protein; the amino acid sequence of the modified spidroin protein is shown in SEQ ID NO.1, and the modified spidroin protein has protease cleavage sites KSS, DSS, ESS and RSS , the prepared suture can gradually dissolve in the body, so the prepared suture does not need to be removed after operation, which can reduce the pain of patients; at the same time, because the modified spidroin protein has good biocompatibility and toughness, the prepared suture has good Mechanical properties and biological qualities. The absorbable medical suture raw material provided by the invention also includes hydroxymethyl chitosan, which can promote wound healing. The present invention also provides a preparation method of absorbable medical suture, specifically: dissolving the raw material of absorbable medical suture to obtain spinning stock solution; drying the spinning stock solution after wet spinning to obtain the absorbable medical suture Medical sutures.

Provided herein are methods of improving solubilization, extraction, and isolation of recombinant spider silk proteins with a salt and alcohol buffer. Provided herein are methods of solubilizing a recombinant spider silk protein from a host cell, comprising: providing a cell culture comprising a host cell, wherein the host cell expresses a recombinant spider silk protein; collecting an insoluble portion derived from the cell culture, wherein the insoluble portion comprises the recombinant spider silk protein; adding the insoluble portion of the host cell to a solution comprising a salt and an alcohol, thereby solubilizing the recombinant spider silk protein in the solution.

The degradable tissue repair material provided by the present invention, the raw material of the degradable tissue repair material includes recombinant spidroin protein; the amino acid sequence of the recombinant spidroin protein is shown in SEQ ID NO.1; the recombinant spidroin protein, By inserting a protease cleavage site between two adjacent spidroin protein domain monomers, when the recombinant spidroin protein is used in the human body, the protease cleavage site on the recombinant spidroin protein can be eliminated by the human body. The enzymatic hydrolysis of the recombinant spidroin protein can make the biomaterials made of the recombinant spidroin protein gradually degrade in the human body, so using the above-mentioned recombinant spidroin protein as raw material to prepare tissue repair materials can ensure the mechanical stability of the original spidroin protein. While ensuring performance or biological quality, it can also guarantee its degradability and meet clinical requirements.

The invention provides a production method of spider-fibroin-implanted silk. The production method of the spider-fibroin-implanted silk comprises the following step: (1) implanting silkworm eggs withspider fibroin mucus from glandular ducts of spiders in vivo so as to obtain spider-fibroin-implanted silkworm eggs; (2) hatching the silkworm eggs so as to obtain young silkworms, namely spider-fibroin-implanted silkworms; (3) feeding the spider-fibroin-implanted silkworms by adopting a silkworm feeding method; and (4) waiting for the spider-fibroin-implanted silkworm to spin silk, thereby obtaining the spider-fibroin-implanted silk. Preferably, before the step (1), the production method of the spider-fibroin-implanted silk further comprises a step (11) of rinsing internal organs of the spiders with normal saline in vivo under a microscope under sterile indoor conditions; and, between the steps (1) and the step (2), the production method of the spider-fibroin-implanted silk still furthercomprises a step (12) of putting the silkworms into a thermal container so as to accelerate spinning of the silk. The invention further provides the spider-fibroin-implanted silk produced by the production method. The production method of the spider-fibroin-implanted silk is capable of easily producing the spider-fibroin-implanted silk; and moreover, the produced spider-fibroin-implanted silk hashigher strength and elasticity compared with common silk, so that needs of new materials for military protection and medical use are met. In addition, the production method of the spider-fibroin-implanted silk is smartly designed, easy to operate, low in cost, and suitable for large-scale application and application.

The invention relates to a preparation method and application of nano-particle functionalized artificial spider silk. The preparation method comprises the steps that nascent artificial spider silk is taken as a template, nano-particles are prepared in situ through chemical reaction, and nano-functionalization of spider silk is realized; specifically, recombined spider silk protein nascent fibers are soaked in an ion solution, and nucleation sites of the nano-particles are formed through ions in combination with protein molecular chains in the nascent fibers in advance; and the artificial spider silk is soaked by adopting a composite ion solution, and in-situ synthesis is carried out on the nano-particles which are uniformly dispersed and uniform in particle size in the artificial spider silk. The preparation method is simple and convenient, the produced composite fiber contains nano-particles which are small in particle size and uniformly dispersed, the mechanical property of the fiber can be further improved through nano functionalization, and the toughness of the fiber exceeds that of natural spider traction fibers; and meanwhile, after multiple times of water washing and drying, the functional properties of the composite fiber are almost not affected, and different kinds of nano-particles can be prepared in the fiber, so that the requirements of people on fiber materials in different scenes are met.

The invention relates to the technical field of biomedical science, and provides spider silk protein bone nails and a preparation method thereof. The preparation method of the spider silk protein bonenails comprises the following steps of (1) preparing a spider silk protein solution; and (2) injecting the spider silk protein solution into a mold, performing treatment with methanol or ethanol, andperforming drying so as to obtain the spider silk protein bone nails. The preparation method of the spider silk protein bone nails provided by the invention is simple to operate and can be in industrialized production; and the prepared spider silk protein bone nails have the advantages of being high in strength, high in tenacity, light, resistant to ultraviolet rays, biodegradable and the like, can be degraded into amino acids and polypeptide in human bodies, and do not have any toxic or harmful effects on the human bodies.