8 results about "Recombinant Insulin" patented technology

The invention relates to a TCEP-based recombinant proinsulin inclusion body denaturation and renaturation method, according to the method, TCEP is added as a reducing agent in the denaturation and renaturation process, compared with the prior art, the yield of proinsulin denaturation and renaturation is increased, and the yield of insulin is remarkably increased. Besides, TCEP is high in reducibility, wide in applicable pH range and extremely low in toxicity, so that the production efficiency and the process stability of the recombinant insulin production process can be remarkably improved, the harm to operators and the environment is reduced, and the requirements of green chemistry and safe production are met.

The application discloses a kind of modular serum-free cell culture medium and its application, belong to biomedical technology field.The culture medium includes: epidermal growth factor, recombinant insulin, cholesterol-phospholipid nanoparticles, beta-mercaptoethanol, pluronic F-68, glucose / glutamine, trace element mixed solution.The modular serum-free cell culture medium provided by the application can not only meet the large-scale culture of stem cells, but also be used for the industrial production of in vitro diagnostic reagent raw materials such as recombinant antigen, viral vector, recombinant protein and monoclonal antibody, with wide cell adaptability, low production cost, significant economy, stable process, and strict requirements of FDA / EMA for biological products.

The application provides a crystallization process and product of recombinant insulin, and relates to the technical field of biological medicine; the crystallization process comprises the following steps: mixing and dissolving recombinant insulin, an organic acid, an organic solvent, a salt and water, adding a zinc substance to obtain a crystallization solution; adjusting pH, and then standing after low-temperature stirring; and separating the recombinant insulin crystal; in the crystallization solution, when the insulin concentration is 5g / L, the organic solvent concentration is 10%-30%, and pH=8.0-9.3; when 5<insulin concentration≤10g / L, the organic solvent concentration is 15%-30%, and pH=8.8-9.5. In the absence of phenolic substances, the application realizes the successful crystallization of high-concentration recombinant insulin by adopting specific concentrations of recombinant insulin and organic solvent and adjusting pH; the crystal form is similar to that in the presence of phenol; and the obtained crystalline recombinant insulin is applied to the preparation of a drug for treating diabetes.

The invention provides a crystallization process of recombinant insulin glargine and a product, and relates to the technical field of biological medicines. The crystallization process comprises the following steps: mixing and dissolving recombinant insulin glargine, organic acid, an organic solvent, salt and water, and adding zinc substances to obtain a crystallization solution; adjusting the pH value, stirring at low temperature, standing, and separating the recombinant insulin glargine crystal; in the crystallization solution, when the concentration of insulin is 5g / L, the concentration of the organic solvent is 10-30%, and the pH is 8.0-9.3; when the concentration of insulin is more than 5 and less than or equal to 10g / L, the concentration of the organic solvent is 15-30%, and the pH is 8.8-9.5. Under the condition of no phenolic substances, the successful crystallization of the high-concentration recombinant insulin glargine is realized by adopting the specific concentration of the recombinant insulin glargine and the concentration of the organic solvent and combining the conditions of pH regulation and the like; the crystal form of the recombinant insulin glargine is similar to that of the insulin glargine with phenol, and the obtained crystallized recombinant insulin glargine is applied to preparation of drugs for treating diabetes.

The present invention relates to a recombinant insulin precursor. To improve the recombinant production of insulin, a recombinant insulin precursor according to the chemical formula LBCA is provided, wherein A is the A-chain of insulin, B is the B-chain of insulin, C is a C-peptide connecting the A-chain and the B-chain, and L is an N-terminal L-peptide. The C-peptide comprises an amino acid sequence according to SEQ ID NO: 1 and / or the L-peptide comprises an amino acid sequence according to SEQ ID NO: 2.

This invention provides a mesenchymal stem cell culture medium, its preparation method, and its application. The mesenchymal stem cell culture medium of this invention includes a basal culture medium and functional additives. The functional additives include: recombinant human albumin, recombinant transferrin, recombinant bone morphogenetic protein; recombinant insulin or insulin-like growth factor, recombinant fibroblast growth factor, recombinant epidermal growth factor, recombinant platelet-derived growth factor; thyroid hormones, ethanolamine, carnitine, glucocorticoid regulators, retinoic acid compounds; heparin sodium, signal-regulating lipids, cholesterol, DL-α-tocopherol acetate, linolenic acid, oleic acid, stearic acid; prolactin F68, and Tween 80. The mesenchymal stem cell culture medium of this invention does not contain immunogenic proteins or blood products derived from animals or humans. All components can be supplied with GMP-compliant raw materials and the process can be controlled, ensuring the controllability, safety, and reproducibility of the culture process.

The invention discloses a serum-free additive E6 for a chicken primordial germ cell culture medium and application of the serum-free additive E6, and belongs to the technical field of animal husbandry and biology. The E6 serum-free additive is composed of six components necessary for in-vitro proliferation of chicken PGCs, and specifically comprises DL-alpha-tocopheryl acetate, BSA (fatty acid-free component V), human recombinant insulin, corticosterone, linolenic acid and triiodothyronine. After the E6 serum-free additive is added into a chicken PGCs culture medium, the basic form of cells can be effectively maintained, long-term in-vitro proliferation of the cells is supported, expression of specific marker genes and proteins such as reproduction and pluripotency is maintained, and meanwhile the long-term subculture capacity and gonad migration potential of the PGCs are guaranteed. The E6 serum-free additive disclosed by the invention has the advantages of simple and clear components, simplicity and convenience in preparation and relatively low cost, and can be widely applied to the fields of poultry germplasm resource preservation, gene editing, biological breeding and the like.

The invention discloses a serum-free culture medium with definite chemical components for human umbilical cord mesenchymal stem cells, and belongs to the technical field of cell culture. The serum-free culture medium comprises a human umbilical cord mesenchymal stem cell basic culture medium and an additive, wherein the additive is prepared from Activin A, GDNF, R-spondin 2, SAG, SC79, human recombinant transferrin, vitamin C, human recombinant insulin and beta-sodium glycerophosphate. The cell culture auxiliary factors in the additive are added to replace serum to be used in a complete culture medium, so that the problems of unstable culture medium batch, easiness in pathogen pollution, high cost and the like caused by undefined serum chemical components are solved. Meanwhile, by adopting the serum-free culture medium disclosed by the invention, the proliferation efficiency can be improved and the cell activity can be better maintained in in-vitro proliferation.