98 results about "Triple helix" patented technology

The invention relates to a recombinant I-type humanized collagen with crosslinking property as well as a preparation method and application thereof, and belongs to the technical field of recombinant proteins. The invention relates to recombinant I-type humanized collagen with crosslinking property. The amino acid sequence of the recombinant I-type humanized collagen comprises an amino acid sequence as shown in SEQ ID No.1 or a derivative amino acid sequence with the sequence identity of more than 90% with the sequence as shown in SEQ ID No.1; the recombinant I-type humanized collagen has a triple helix structure. A complete triple helix structure can be efficiently expressed, and the hydrogel has efficient cross-linking capability, so that the hydrogel can be efficiently and stably formed through cross-linking.

The present invention relates to the fields of medicine and cosmetology, and discloses a cell active peptide composition that promotes the regeneration of multi-type collagen structural proteins, its preparation method, and its application in medicine and cosmetology. The composition uses raw materials such as tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, γ-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, and palmitoyl pentapeptide-4. The composition can promote the coordinated expression of skin type I, type III, type IV, type VII, type XVII collagen, fibronectin, elastin, and glycosaminoglycans; prevent collagen cross-linking, maintain triple helix structure, and inhibit collagen degradation. Applied to dermatology and cosmetology, it can accelerate the regeneration of structural proteins such as skin collagen, repair skin damage, reduce wrinkles such as crow's feet, tear groove lines, nasolabial folds, and under-eye lines, solve the skin aging problem caused by the loss of structural proteins such as collagen, and improve skin elasticity and firmness.

The invention belongs to the technical field of biological medicine, and particularly relates to recombinant human XVII micromolecular collagen as well as a preparation method and application thereof. According to the invention, a recombinant human XVII type micromolecular collagen (Re-Hcol17) is constructed, and a stable triple helix structure is formed by connecting the 15th collagen domain, the 12th collagen domain, the 5th collagen domain and the 1st collagen domain in series. Experimental results prove that the Re-Hcol17 can significantly promote NIH / 3T3 fibroblast proliferation within the concentration range of 10-100 [mu] g / mL, the effect is better than that of natural human collagen, and extracellular matrix adhesion can be effectively promoted. Through genetic engineering and fermentation process innovation, the bottleneck that the human-derived XVII type collagen is difficult to efficiently express and unstable in structure is broken through, and a foundation is laid for wide application of the human-derived XVII type collagen in the fields of biological medicines and cosmetics.

The present invention relates to the technical field of protein engineering and provides a recombinant humanized type III collagen having a triple helix structure, a preparation method therefor, and a use thereof. The amino acid sequence of the recombinant humanized type III collagen comprises n core units, wherein n is 1 or an integer greater than 1, and the amino acid sequence of the core unit is as shown in SEQ ID NO: 1. The present invention further provides a method for constructing an expression system and a method for preparing a recombinant humanized type III collagen. The present invention further provides an actual industrial use on the basis of the characteristics of the protein. The recombinant humanized type III collagen provided by the present invention has high purity, good stability, and high biological activity. Experiments have shown that the recombinant humanized type III collagen has various biological activities such as promoting cell proliferation and migration.

The invention relates to the technical field of synthetic biology, in particular to polypeptide with a triple helix structure, recombinant XII type humanized collagen and application of the recombinant XII type humanized collagen. The recombinant XII type humanized collagen is successfully expressed and prepared, has a triple-helix structure and good biological activity including promotion of cell proliferation activity, cell adhesion activity and inhibition of MMP-1, is used as a biological material derived from a human body, does not generate immunological rejection and anaphylactic reaction when applied to the human body, and has a good application prospect. The composition can be used for medical or non-medical application of a plurality of tissues and organs of a human body, filling, compatibilizing or repairing, and promotion of skin compactness or wrinkle resistance and other scenes.

The invention provides a medical-grade recombinant three-type collagen and application thereof. The recombinant humanized three-type collagen has certain cell proliferation promoting capacity and is free of cytotoxicity. The medical-grade recombinant three-type collagen provided by the invention not only has the capability of promoting cell proliferation, is free of cytotoxicity and high in cell adhesion activity, but also has a three-helix structure characterized by the collagen and is extremely high in stability, the purity can reach about 85% after the collagen is induced for 120H in a 100L fermentation tank, the difficulty of a downstream purification process is greatly reduced, and the production cost is reduced. The effect in the aspect of cell activity is obvious.

The invention specifically discloses a recombinant XVII type collagen and a preparation method thereof, and relates to the technical field of genetic engineering. The invention provides a recombinant XVII type collagen protein and a preparation method of the recombinant XVII type collagen protein. Compared with the prior art, the recombinant XVII type collagen pichia pastoris engineering strain provided by the invention is high in yield, the protein is not easy to degrade, and the recombinant XVII type collagen pichia pastoris engineering strain has a stable triple helix structure.

The invention discloses recombinant collagen III + X as well as a preparation method and application thereof, and belongs to the technical field of biochemical engineering. The amino acid sequence of the recombinant collagen III + X provided by the invention is shown as SEQ ID NO: 1, and the recombinant collagen III + X is formed by repeating human III type collagen 387-467 amino acid fragments twice and connecting human X type collagen 521-680 amino acid fragments to the C end. Experiments prove that the recombinant III + X collagen and proline hydroxylase are co-expressed to form a stable triple helix structure, and the recombinant III + X collagen has great advantages in the aspects of cell migration, safety and the like, and can be used as a raw material for preparing wound dressings and skin care products.

Methods of providing biologically synthesized human structural material Type VI collagen are provided. The collagen of the present application comprises an amino acid sequence set forth in SEQ ID NO: 2 or a variant amino acid sequence that is mutated from the amino acid sequence set forth in SEQ ID NO: 2, the variant amino acid sequence retaining the function of the amino acid sequence set forth in SEQ ID NO: 2. The collagen of the present application is capable of promoting cell adhesion and has a triple helix structure.

The invention discloses a high-biological-activity human-derived I-type collagen variant, a composite material based on the high-biological-activity human-derived I-type collagen variant and a preparation method. According to the invention, the COL109 fragment which can be efficiently secreted and expressed in pichia pastoris and has a triple helix structure is obtained through screening; an adhesion domain (GPP) 10 is further fused to improve the mechanical strength, and a variant COL109 (GPP) 10 is obtained; and finally, compounding with polycaprolactone (PCL) according to an optimized ratio (1: 1) to obtain the composite material with high biological activity (promoting osteogenic differentiation and wound healing) and excellent mechanical properties. The material is suitable for bone tissue engineering scaffolds, skin regeneration dressings and the like.

The application discloses a recombinant collagen type XVII and a preparation method thereof, and relates to the technical field of genetic engineering. The application provides a recombinant collagen type XVII. Compared with the prior art, the recombinant collagen type XVII provided by the application has high yield of Pichia pastoris engineering bacteria, is not easy to be degraded, and has a stable triple helix structure.

The invention discloses a bone repair material as well as a preparation method and application thereof. The bone repair material is prepared from recombinant human collagen, recombinant human bone morphogenetic protein 2 and hydroxyapatite, wherein the ratio of the inorganic matter to the organic matter in the bone repair material is (64-66): (36-34). The bone repair material disclosed by the invention takes the recombinant human collagen as a core raw material, so that the components and the structure of the prepared material are ensured to be highly consistent with those of natural bone tissues; secondly, by adding the recombinant human bone morphogenetic protein 2, the osteogenic activity of the material is further enhanced. The finally obtained recombinant human collagen active bone repair material completely retains the triple helix structure of the recombinant human collagen, so that the recombinant human collagen active bone repair material has complete biological activity, and also has the outstanding advantages of good osteogenic activity, no disease and virus transmission risk and no immunogenicity.

The present disclosure provides recombinant type XVII collagen having a triple helix structure and uses thereof, which belong to the field of protein engineering technology. The recombinant type XVII collagen has an amino acid sequence formed by tandem repeats of n or more core units, where n is an integer of 1 or more, and the core unit has the amino acid sequence shown in SEQ ID NO: 1. The present disclosure also provides methods for constructing multiple expression systems and methods for preparing recombinant type XVII collagen. provide.

The invention provides recombinant II-type collagen as well as a preparation method and application thereof, and belongs to the technical field of bioengineering. The recombinant II-type collagen monomer is obtained by splicing Gly-X-Y repetitive gene sequences of multiple sections of triple helix regions in human II-type collagen, immunodominant epitopes, unstable amino acid sequence 3 conjunctions and glycosylation sites are avoided during selection, then the recombinant II-type collagen monomer is repeated for multiple times, and the recombinant II-type collagen monomer is obtained. The recombinant II type collagen with the molecular weight of 40-50 kDa is obtained; the recombinant II-type collagen is beneficial to expression of pichia pastoris, has the effects of promoting adhesion and migration of cartilage cells, can also promote differentiation into cartilage cells, and has good practicability.

This invention relates to the field of biomedical materials, providing non-denatured type II collagen ultrafine powder with cartilage tissue-targeted growth and structural repair functions, its bioactivity preservation preparation method, and its application in joints. The ultrafine powder has an average particle size of 0.5-50 μm, a triple helix structure integrity rate of not less than 85%, a thermal denaturation temperature of 38-48℃, and retains chondrocyte binding domains CB10, CB11, and MMP cleavage sites. The preparation method includes raw material pretreatment, time-limited enzymatic hydrolysis, graded purification, and ultra-low temperature ball milling. The product can be used for the repair and treatment of cartilage degenerative diseases such as osteoarthritis and cartilage damage.

The invention discloses a polypeptide, a recombinant XI type humanized collagen formed by the polypeptide and application of the recombinant XI type humanized collagen. Through a large number of screening studies, it is found that a natural human XI type collagen core functional region of recombinant XI type humanized collagen with a triple helix structure can be formed in the in-vitro biosynthesis process. On the basis, the recombinant humanized polypeptide is designed and prepared on the basis of a natural XI type collagen core functional area, and the polypeptide can form a stable triple-helix structure. The recombinant XI type humanized collagen provided by the invention has good biological activities, including promotion of cell proliferation activity, promotion of cell adhesion activity and promotion of extracellular matrix generation or reconstruction, and is suitable for various medical or non-medical applications.

The invention belongs to the technical field of recombinant collagen sequence analysis, and discloses a recombinant collagen sequence analysis method based on AI deep learning. Comprising the following steps: extracting a natural III-type collagen sequence; applying the natural III-type collagen sequence to the trained model to generate a thermodynamic diagram; generating an optimized natural III type collagen sequence based on the thermodynamic diagram; performing fine tuning on the AlphaFold2 model by using the fine tuning data set, and obtaining an integrated structure model by introducing spiral torsion constraint and hydroxyproline simulation parameters; loading the optimized natural III-type collagen sequence, and predicting the stability of the triple helix structure of the sequence through the structure model; and extracting a natural III-type collagen sequence which passes the stability detection, and outputting a report file by using a skin permeability prediction model to realize recombinant collagen sequence analysis based on AI deep learning.

The invention discloses application of a triple-helix structural element in RNA (Ribonucleic Acid) drug design, and belongs to the field of biological medicines. According to the application, an ENE element or a mutant of the ENE element is used as a nucleic acid stabilizing element, and the interaction between 3 'UTR and poly (A) is optimized, so that the mRNA stability is enhanced, and the protein translation efficiency is further improved, and therefore, the problem of low protein expression quantity in RNA therapy is effectively solved. The mRNA sequence optimization strategy provided by the invention provides a new method for vaccine design in the field of infectious disease and tumor immunotherapy, and has a wide application prospect.

The invention belongs to the technical field of gene recombination, and particularly relates to a recombinant human III-type triple helix collagen, a recombinant yeast strain for expressing the recombinant human III-type triple helix collagen and a construction method of the recombinant human III-type triple helix collagen. The amino acid sequence of the recombinant human III-type triple-helix collagen is shown as SEQ ID NO. 1, and the recombinant human III-type triple-helix collagen has triple-helix conformation close to a natural biological structure of collagen and has good application value and potential. According to the recombinant yeast strain disclosed by the invention, human-derived III-type collagen and proline hydroxylase are co-expressed in yeast cells without YPS1 protease, and meanwhile, a gene beneficial to vesicle transport and protein folding is transferred, so that hydroxylation of the recombinant collagen can be realized, a natural triple helix conformation is formed, and the recombinant yeast strain can be used for preparing a recombinant collagen product. The degradation problem of the collagen in the expression process can be solved, and the extracellular secretion expression of the triple helix collagen is realized.

The present disclosure provides a recombinant humanized collagen VIII, which is a triple helix structure, has good cell adhesion activity, and is applied to human body without immune rejection and allergic reaction.

This invention discloses a method for maintaining the triple helix structure during the preparation of methacrylated recombinant collagen hydrogels, belonging to the field of genetic engineering technology. This invention optimizes the reaction parameters of the methacrylic anhydride (MA) chemical cross-linking process of recombinant collagen. Through screening, the titration method with an addition rate of 2 μL / min for MA was selected. The reaction temperature and the amount of MA added were optimized, and it was found that a MA:Col ratio of 1:2 and a reaction temperature of 35℃ resulted in good triple helix maintenance. This invention, through the titration method of adding methacrylic anhydride, greatly maintains the triple helix structure of recombinant collagen during the preparation of hydrogels, preserving excellent biocompatibility and showing broad application prospects in the field of biomaterials.

The invention provides a method for preparing recombinant human collagen. The method comprises the following steps: carrying out fermentation culture treatment on engineering bacteria, expressing pre-collagen by the engineering bacteria, and carrying out purification treatment on a fermentation culture treatment product, so as to obtain the recombinant human collagen, wherein the fermentation culture treatment is three-stage fermentation, and the final liquid loading amount of the three-stage fermentation is not less than 9 tons. The method can be used for preparing the recombinant human collagen with a complete triple helix structure, low immunogenicity, high safety and high purity.

A self-crosslinking recombinant humanized collagen polymer biomaterial and a preparation method therefor are provided. A collagen self-assembled element includes (i) at least 21 consecutive amino acids at the C-terminus of the amino acid sequence as shown in SEQ ID NO: 1; or an amino acid sequence that is obtained by means of substitution, deletion or addition of one or more amino acids in the amino acid sequence as described in (i) and that retains the function of promoting the self-crosslinking of recombinant type III humanized collagen to form a triple helix structure; or an amino acid sequence that has at least 90%, 92%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence as described in (i) and that retains the function of promoting the self-crosslinking of recombinant type III humanized collagen to form a triple helix structure.

The invention discloses triple helix recombinant collagen as well as a preparation method and application thereof, and belongs to the technical field of gene engineering and protein engineering. According to the invention, on the basis of the amino acid sequence of natural collagen, the amino acid sequence is designed by means of bioinformatics and the like, a novel triple-helix recombinant collagen is constructed, and the triple-helix recombinant collagen has a good triple-helix structure, biocompatibility and biological activity, and can be used for preparing the collagen. The proliferation, the adhesion and the migration of the L929 cells can be obviously promoted, and the method has a wide application prospect in various fields such as food, medicines, cosmetics or instrument products.

The invention belongs to the field of protein engineering. The invention provides high-stability non-hydroxylase modified recombinant humanized collagen as well as a preparation method and application thereof. The amino acid sequence of the high-stability non-hydroxylase modified recombinant humanized collagen comprises one of amino acid sequences shown as SEQ ID NO.1 or SEQ ID NO.2. The invention further provides a preparation method of the high-stability non-hydroxylase modified recombinant humanized collagen. Under the condition that the recombinant humanized collagen does not need to be subjected to hydroxylation modification, the triple helix stability is remarkably improved, a streptococcus Scl2 collagen-like protein triple helix stable structural domain is excavated through molecular dynamics simulation to be embedded with a natural collagen sequence, the natural sequence is reserved, the biological activity of the collagen is ensured, and the recombinant humanized collagen has the advantages of being simple in structure, convenient to use and the like. The dependence of a traditional recombinant expression technology on hydroxylation modification of proline is also avoided.

A new preparation method of recombinant humanized collagen hydrogel is provided. The method comprises the following steps: reacting recombinant humanized collagen and a dialdehyde cross-linking agent, wherein the concentration of the dialdehyde cross-linking agent is 0.01-0.25%, and the recombinant humanized collagen is one or more of recombinant humanized type II, type III and type XVII collagen, and the recombinant humanized collagen has a triple helix structure. The present application determines the aldehyde cross-linking agent suitable for recombinant humanized collagen, especially the recombinant humanized collagen with a triple helix structure and the optimum reaction conditions.

The invention belongs to the technical field of biological medicines, and particularly relates to a tiny binding protein of a targeted vascular endothelial growth factor receptor 2 (VEGFR2) as well as a construction method and application of the tiny binding protein. The amino acid sequence of the tiny binding protein is shown as SEQ ID NO. 5, and the tiny binding protein is of a triple helix structure. The tiny binding protein of the vascular endothelial growth factor receptor 2 (VEGFR2), provided by the invention, can be used as a tool protein to be combined with VEGFR2 protein and is used for regulating vascular endothelial growth.

This invention relates to bioengineering, specifically disclosing a method for preparing recombinant humanized type III collagen with a triple-helix structure, comprising the following steps: S1, constructing a ppICZαA recombinant expression plasmid containing a human type III collagen fragment, and transforming the recombinant expression plasmid into yeast; S2, screening high-copy transformant strains from the strain and performing shake-flask level induction expression tests; S3, conducting high-density fermentation in a fermenter to obtain a large amount of secretible target protein. This invention, by constructing a ppICZαA recombinant expression plasmid containing a human type III collagen fragment and electroporating it into a yeast expression system, screens for yeast strains capable of expressing recombinant humanized type III collagen fragments, and conducts high-density fermentation in a fermenter, followed by purification after fermentation, yields pure recombinant collagen with a triple-helix structure, which can be applied in skincare products, functional skincare products, medical devices, and biomedical materials.

Provided is a method for preparing a recombinant human collagen. The method comprises: constructing a genetically engineered bacterium which expresses a prolyl hydroxylase and a recombinant human collagen; culturing the genetically engineered bacterium and performing digestion treatment by means of pepsin to obtain a recombinant human collagen type I or type III. The prolyl hydroxylase comprises P4Ha2, P4Hb and vP4H, and by means of controlling the ratio of the copy numbers of the genes thereof, the recombinant human collagen and a natural human collagen are consistent in hydroxylation modification. Also provided is a large-scale production method for the recombinant human collagen. The prepared recombinant human collagen has completely the same sequence composition and length and substantially the same hydroxyproline content as the natural human collagen, and also has a stable full-length triple helix structure and biological activity without immunogenicity and potential risk of viruses.

The application discloses a pyridaben lateral flow analysis method and test paper based on triple helix bivalent aptamer and nano enzyme, and belongs to the fields of analytical chemistry, medicine, environment, food safety detection and nano biosensing. By constructing a triple helix bivalent aptamer, the binding rate of the aptamer and the target is greatly improved, and the response time is shortened to 1 / 6 of that of a conventional aptamer. With the excellent peroxidase-like activity of the Au@Pt nano enzyme, the sensitivity of the Apt-LFA test for detecting pyridaben is improved by 5 times. The streptavidin and streptavidin-biotin-DNAc sprayed in the detection zone and the control zone do not need to be changed, and only the nucleic acid chain part on the probe Au@Pt NPs@poly-DNA needs to be changed to detect another substance. A new lateral chromatography method for rapid, sensitive and low-cost detection is developed, and the nano enzyme triple helix aptamer test paper has great application potential.