176 results about "Recombinant Human Epidermal Growth Factor" patented technology

The invention discloses a hydrogel dressing containing a recombinant human epidermal growth factor. The hydrogel dressing consists of 100 volume parts of mixed solution A and 2 to 8 volume parts of cross-linking agent; the mixed solution A is prepared by mixing 2.0 to 5.0 mass percent of polyvinyl alcohol aqueous solution and 0.5 to 2.0 mass percent of sodium alginate aqueous solution according to the mass ratio of (2-5): 1, and adding 0 to 0.1 mass percent of sodium benzoate, 5 to 20ug/g of recombinant human epidermal growth factor and 2.0 to 10 mass percent of glycerol; and the gel dressing is prepared by taking the 2 to 8 volume parts of cross-linking agent prepared by mixing sodium borate solution and calcium chloride solution, dripping the cross-linking agent into the 100 volume parts of mixed solution A, standing and defoaming to obtain the gel dressing. The dressing has good biocompatibility and mild and simple preparation condition, can be formed at room temperature, can effectively keep the activity of the recombinant epidermal growth factor and has controlled release effect on the recombinant epidermal growth factor. The dressing has good effect of treating the wound of rats suffering from diabetes mellitus, and can accelerate healing of the wound.

This invention includes signal peptide gene phoAspL10, expression plasmid pBL10EGF, host cell of bacillus coli BL21 (DE3) inverted by expression plasmid pBL10EGF, engineering strain BE-2 screened from the host cell and preparation method of rhEGF. The strain BE-2 can excrete and express rhEGF directly into culture after fermentation and the quantity is 384mg/l.The purity is more than 95% and specific activity more than 1.0*106IU/mg protein. The product rhEGF reserves crude space configuration and shows high biological activity.

The invention relates to an external gel used for treating diabetic foot, and a preparation method and an application thereof. Specifically, the invention relates to an external gel preparation with oxygen-carrying and antibacterial materials, and containing recombinant human epidermal growth factor (rhEGF). The preparation basically comprises: a medicine component recombinant human epidermal growth factor (rhEGF), an oxygen-carrying material, an antibacterial material, a gel substrate, a stabilizing agent, a humectant, a penetration enhancing agent, a surfactant, and a pH regulation agent.

The invention relates to an external composition, and a preparation and an application thereof and belongs to the field of medicines. The external composition provided by the invention is a medicine composition which contains chlorhexidine or officinal salt thereof, or benzalkonium bromide or officinal salt thereof and a recombinant human epidermal growth factor or a recombinant cattle alkali fibroblast growth factor. According to the invention, the medicine composition is also further prepared into gel, liniment, ointment or paste, and the gel further comprises chitosan or officinal salt thereof and hyaluronic acid or officinal salt thereof. When being used for treating vagina inflammation, the external composition disclosed by the invention can improve a vagina microenvironment, promote growth of granulation tissue and repair vagina micro damage; and when being used for skin wound atraumatic restorative treatment, the external composition also can speed up restoration and regeneration of a wound tissue, promote healing and reduce scar formation, so that the external composition has a wide medical application prospect.

The invention relates to the technical field of gene recombination, in particular to a method for preparing soluble and high-activity recombinant human epidermal growth factor; the invention solves the problems that the human epidermal growth factor which is obtained by using the chemical synthesis method and the biological extraction method which have different self-defects respectively and by using the gene recombination method has lower purify and low yield and activity, and comprises the following steps of: constructing the gene cloning of the human epidermal growth factor, replacing codons which are not beneficial to be expressed by Escherichia coli to obtain optimized gene sequence, adding molecular chaperone of PDI to help target gene to be expressed effectively in Escherichia coli, and embedding HIS label and enzyme cutting site of protein in the sequence to help the target gene to be purified effectively, and the identification of biological activity; the obtained human epidermal growth factor has high purify, high yield, high activity, low cost, simple production technique and stable product quality; the product can be used for manufacturing drugs or articles for beauty and cosmetics and is beneficial to be promoted to the public.

The invention discloses wrinkle removing, anti-aging, skin activating and scar fading repair essence which comprises components in parts by weight as follows: 600-850 parts of sterile water, 60-200 parts of glycerin, 30-80 parts of sodium pyrrolidone carboxylic acid, 30-80 parts of 1,3-butanediol, 3-10 parts of collagen, 5-20 parts of panthenol, 3-10 parts of allantoin, 10-40 parts of hydroxyproline, 2-10 parts of bisabolol, 2-10 parts of hyaluronic acid, 0.5-2 parts of recombinant human epidermal growth factors and 1-5 parts of diazolidinyl urea. The preparation method comprises the following steps: (1), sterile water, glycerin, sodium pyrrolidone carboxylic acid, 1,3-butanediol and collagen are mixed uniformly with allantoin and are heated for 20 min at 85 DEG C; (2), the mixture is stirred for 20 min at the speed of 30 rpm and is cooled; (3), panthenol and hydroxyproline are added when the temperature is 55 DEG C, the mixture is uniformly mixed, residual components are added when the temperature is 40 DEG C, the mixture is uniformly mixed and then homogenized for 30s, and discharging is performed when the temperature is 30 DEG C, and the repair essence is obtained.

The invention relates to the technical field of cell culture, and in particular to a serum-free culture medium, a preparation method thereof and a culture method of mesenchymal stem cells. The culturemedium consists of the following components: non-essential amino acid, glutamine, hydrocortisone, dexamethasone, polyvinyl alcohol, recombinant human insulin, recombinant human transferrin, a recombinant human epidermal growth factor, a recombinant human basic fibroblast growth factor, a recombinant human Wnt-3a protein, a recombinant human fibronectin, recombinant human laminin, L-glutathione, L-ascorbic acid, beta-mercaptoethanol, ethanolamine, Pluronic F-68, Tween 80, cholesterol, adenine, sodium selenite and a basic culture medium. According to the invention, polyvinyl alcohol can replaceserum albumin in a serum-free culture medium in carrier function, so that proliferation capacity of mesenchymal stem cells is effectively improved. Polyvinyl alcohol has more stable performance thanserum albumin, and the problem that proliferation capacity and passage capacity of stem cells are reduced can be effectively solved.

The invention relates to a method for producing, purifying and appraising protein, in particular to a method for expression of recombinant human her2/neu ICD in Pasteur yeast as well as optimized large scale industrialized fermentation production and purification of the recombinant human her2/neu ICD.

The invention provides a serum-free culture medium used for full-suspension culture of MDCK (Madin Darby Canine Kidney) cells. The culture medium is prepared from a DMEM/F12 culture medium containing 5g/L of NaCl and the following components: sodium stannite, recombinant human epidermal growth factors, hydrocortisone, recombinant full-chain insulin, prostaglandin E1, human transferrin, thyroxine (T3), vitamin E, cholesterol, ethanol amine, beta-mercaptoethanol, Tween-80, Hypep1510, recombinant human serum albumin ACF and mannitol. With the adoption of the culture medium provided by the invention, the full-suspension culture of the MDCK cells can be carried out very well under the condition that serum is not added; after the MDCK cells are continuously cultured for 20 generations, the average proliferation concentration of the MDCK cells is 2.308*10<6>/mL, the average cell viability is 97.4 percent and the average doubling time is 34.48 hours, so that the serum-free culture medium provides technical supports for developing influenza vaccines of cell matrixes of mammals in China.

The invention relates to a neural stem cells medium and a method for performing human neural stem cells in-vitro long-term culture and amplification by using the neural stem cells medium. The neural stem cells medium comprises the following ingredients by weight proportion: 100-1000 micrograms of heparin sodium, 10-100 micrograms of vitamin E, 5-50 milligrams of insulin human recombinant, 0.5-5 milligrams of putrescine, 2-10 micrograms of sodium selenite, 2-10 milligrams of human transferrin, 2-10 micrograms of progestin, 300 milligrams of L-glutamine, 5.9 grams of 2-[4-(2-Hydroxyethyl)-1-piperazine]ethanesulfonic acid, 10-100 micrograms of recombinant human epidermal growth factors, 10-100 micrograms of recombinant human basic fibroblast growth factors, 20-200 milligrams of vitamin C glucoside and 40,000-400,000 IU (international unit) of gentamicin. By the neural stem cells medium, the technical problems that human neural stem cells are easy to differentiate when cultured in vitro and long-term culture and amplification are difficult to implement are solved.

The invention provides a serum-free medium for liver cancer stem cells and a culture method for the serum-free medium. Based on a DMEM/F12+GlutaMAXTM<-1> medium, the serum-free medium comprises a recombinant human epidermal growth factor, a recombinant human basic fibroblast growth factor-10, an insulin No.1 growth factor, a serum-free complementary nutritional ingredient, heparin and a penicillin and streptomycin double-resistant solution. The serum-free medium has the effects as follows: 1, three special growth factors are added to the serum-free medium in a corresponding proportion; direct serum-free culture is realized, so that liver cancer cells can be pelletized, the pelletizing rate is high, and continuous and stable passage can be realized; 2, the growth factors in the serum-free medium keep a small group of cells in an undifferentiated state; the serum-free medium and the culture method have wide prospect in fundamental research and clinical application of serum-free culture of the liver cancer stem cells and provide rich experiment materials for research of the liver cancer stem cells.

The invention discloses a separation and cultivation method of human adipose stem cells. The method comprises the following steps: first, digesting a fat suction matter by using a Liberase digestive enzyme solution, neutralizing digestive enzyme after digestion is ended, centrifuging, filtering, and further removing parenchyma cells by using a lymphocyte separation solution so as to obtain the human adipose stem cells; second, cultivating the human adipose stem cells by using a serum-free medium, wherein following components are added in the serum-free medium: a recombinant human epidermal growth factor, a recombinant human basic fibroblast growth factor, a recombinant human transforming growth factor-beta, a recombinant human platelet-derived growth factor-BB, a recombinant human stem cell factor, reduced glutathione, coenzyme A, biotin, MEM vitamin solution, MEM amino acid solution, MEM non-essential amino acid solution, a GlutaMAX additive, insulin-transferrin-selenium solution andgentamicin. The method can remarkably increase the yield of adipose stem cells and improve the activity of the adipose stem cells, so as to obtain more adipose stem cells with proliferation ability.

The present invention provides a construction and screening method for recombinant human epidermal growth factor (hEGF) engineering bacteria. The construction and screening method comprises the following main steps: 1, adopting a PCR technology to obtain kanamycin resistance gene, and adopting a chemical synthesis method to obtain an escherichia coli cspA gene function element; 2, respectively optimizing gene sequences encoding SUMO protein and hEGF, and then adopting chemical synthesis to obtain a large amount of expressed fusion protein gene sequences OPT-SUMO-hEGF; 3, linking the fragments obtained from the previous two steps into pET21 plasmid to obtain pET-Kan-cspA-SE plasmid; 4, adopting the pET-Kan-cspA-SE plasmid as a template to construct a transposition body; 5, transforming the transposition body into escherichia coli Origami 2 (DE3); and 6, screening a strain having high fusion protein SUMO-hEGF expression. According to the present invention, genome of the obtained strain is stable, a soluble rate of the expressed hEGF is high, and the method is suitable for large-scale industrial production.

The invention discloses a human umbilical cord mesenchymal stem cell serum-free culture medium and a preparation method thereof, and an acquisition method of a human umbilical cord mesenchymal stem cell serum-free culture solution. The culture medium comprises a basic culture medium and a serum-free culture supernatant concentrated solution, and also comprises the following components and concentrations: 5-100 ng/ml of a vitamin composition, 50-200 [mu]g/ml of grape polyphenol, 1-50 ng/ml of a recombinant human epidermal growth factor, 1-50 ng/ml of a recombinant human basic fibroblast growthfactor, 1-10 ng/ml of a recombinant platelet-derived growth factor-C, 1-10 ng/ml of a recombinant human insulin-like growth factor-1, 50-200 [mu]M of beta-mercaptoethanol, 1-100 [mu]g/ml of human serum albumin, 1-100 [mu]g/ml of glutathione, and 1.2-3.7 g/L of sodium bicarbonate. The human umbilical cord mesenchymal stem cell serum-free culture medium is used for replacing a traditional culture medium containing animal serum to culture human umbilical cord mesenchymal stem cells, and unsafe factors such as human immune response caused by stem cell pollution by pathogens or animal-derived components and heterologous protein are avoided.

The invention discloses a wrinkle-removing, anti-ageing, skin-activating, restoring and whitening composition. The composition comprises the following components in percentage by weight: 15%-30% of hyaluronic acid, 25%-40% of collagen, 0.0001%-0.0009% of recombinant human epidermal growth factor, and 5%-20% of glycerol. Collagen can effectively inhibit generation of melanin and promote metabolism of skin tissue; hyaluronic acid can effectively nourish and moisturize skin, remove wrinkles and beautify looks; recombinant human epidermal growth factor can quicken dropping of senescent cells of a human body and promote reverse differentiation of the cells to generate young cells, can promote regeneration of subcutaneous collagen and recombination of fibrous tissue, improve skin elasticity and reduce skin sensitivity, and can make thick, flushing and expanding facial capillaries atrophy and get thinner and shallower, avoid pigmentation and remove facial toxin residues so as to achieve the efficacy of removing freckles, acne marks and wrinkles as well as tendering and whitening.

The invention aims to provide a simple and inexpensive preparation method for an Ssp intein system mediated recombinant human epidermal growth factor. According to the method, the conventional column cutting and chitin affinity chromatography are replaced with liquid phase cutting and ion exchange chromatography. The column cutting adopted in the conventional Ssp intein system mediated protein purification method is replaced with the liquid phase cutting, so that the loss of the target protein caused by the automatic splitting of fusion protein is reduced; and the expensive chitin affinity chromatography is replaced with the ion exchange chromatography, so that the purification cost of the recombinant human epidermal growth factor is greatly reduced, the operation steps are simplified, and the bottleneck of the large-scale industrial production of the recombinant human epidermal growth factor is overcome.

The invention discloses an umbilical cord mesenchymal stem cell serum-free cultivation method, comprising the following steps: (1) preparing coating buffer for fibronectin, a recombinant human epidermal growth factor and a recombinant human fibroblast growth factor, and coating a plastic culture bottle; (2) taking human umbilical cord tissue Whartonps jelly, cutting into pieces, and adding into the culture bottle in the step (1); (3) for a product obtained in the step (2), adding into a serum-free culture medium; (4) putting into a 5-percent carbon dioxide incubator at 37 DEG C for culture, wherein mesenchymal stem cells overgrow in 10-12 days; (5) washing by a phosphate buffer solution, and performing Triple enzymic digestion; (6) adding the phosphate buffer solution, and stopping the digestion; (7) collecting mesenchymal stem cell supernate; (8) centrifuging; (9) collecting sediments to obtain the needed mesenchymal stem cells. By adopting the method, the mesenchymal stem cells can go down to the 20th generation, the characteristics of the mesenchymal stem cells are kept, diseases can be treated, and application prospect is achieved.

The invention specifically discloses a composition capable of promoting differentiation of autologous preadipocytes and formation of lipid droplets and a preparation method thereof. A commiphora mukul resin extract and a recombinant human epidermal growth factor (rhEGF) are main components of the composition; and the content of the components are that the commiphora mukul resin extract is 1.0 to 3.0% and the recombinant human epidermal growth factor (rhEGF) accounts for 0.01 to 0.10%. According to the invention, the commiphora mukul resin extract and the recombinant human epidermal growth factor are organically combined together; on the one hand, differentiation of autologous preadipocytes is promoted by the recombinant human epidermal growth factor, on the other hand, activity of lipase in decomposition of fat is inhibited by the commiphora mukul resin extract, so formation and accumulation of lipid droplets are promoted, and thus, the volume of a breast can be effectively increased.

The invention relates to a recombinant human epidermal growth factor cationic liposome and a preparation method thereof. The recombinant human epidermal growth factor cationic liposome comprises a recombinant human epidermal growth factor, a cationic liposome and a growth factor protective agent, wherein a part of the recombinant human epidermal growth factor is encapsulated by the cationic liposome. The preparation method comprises the following steps of: step 1. dissolving a panniculus adiposus material and a cationic additive by an absolute ethyl alcohol, carrying out rotary evaporation to get rid of the alcohol to form uniform panniculus adiposus, and adding a citrate buffer solution and hydrating to obtain a crude liposome suspension liquid; step 2. carrying out ultrasonic dispersion to process the crude liposome suspension liquid, and straightening particles by microfiltration membranes to obtain the blank liposome; and step 3. adding the blank liposome solution after mixing a recombinant human epidermal growth factor solution with a growth factor protective agent solution, adjusting pH to 6.0-8.0, and carrying out incubation processing to obtain the recombinant human epidermal growth factor cationic liposome. According to the preparation method, the human epidermal growth factor can be encapsulated by the cationic liposome so that the stability of rhEGF, the intermiscibility of tissues and the affinity of cells can be increased.

The invention relates to a gel, in particular to an induced gel used for repairing skin wound surfaces or cervical mucosa, which not only can solve the therapy problems of skin wounds, cuts and ulcer wound surfaces but also can improve the micro-ecological environments of vaginae and repair the microdamage of the vaginae and meanwhile has good therapeutic effect on cervical erosion. The induced gel comprises Carbomer, triclosan and a recombination human epidermal growth factor, wherein the concentration of Carbomer is 3 to 5 percent, the concentration of triclosan is 0.1 to 0.6 percent, and the concentration of the recombination human epidermal growth factor is 0.00001 to 0.0005 percent. The induced gel can improve the micro-ecological environments of the vaginae and repair the microdamage of the vaginae, thereby being suitable for various vagina inflammations and cervicitis caused by vagina micro-ecological disorder and vagina damage and being suitable for the auxiliary therapy after the clinical therapy on vaginitis and cervicitis and the therapy on cervical erosion; the induced gel also can be used for treating the skin wounds, the cuts and the skin wound surfaces, thereby accelerating the repair and the regeneration of wound tissues, promoting healing and reducing scar formation.

The invention discloses a serum-free umbilical cord mesenchymal stem cell culture medium. The culture medium is used for culturing umbilical cord mesenchymal stem cells and comprises a serum-free basic culture medium and additive components, wherein the addition components include a recombinant human fibroblast growth factor (hFGF), a recombinant human epidermal growth factor (hEGF), insulin hI, L-glutamine, 2-mercaptoethanol, sodium selenite, transferrin, human serum albumin, fibronectin, astragalus polysaccharide APS, IL-3, IL-6, IL-r, a ginseng extract, a rose extract, a jasmine flower extract and vitamin C.

A synergistic pharmaceutical composition for the preparation of topical formulations for use in prophylaxis and treatment of wounds, burn wounds, skin grafts, pressure ulcers, diabetic foot ulcers and other skin diseases is provided. The composition may include one or more synergistically active ingredients and one or more inactive ingredients. The synergistically active ingredients may include Recombinant Human Epidermal Growth Factor (rh-EGF) (REGEN-D™ of Bharat Biotech International Limited) and/or Platelet Derived Growth Factor (rh-PDGF-BB), silver sulfadiazine (SSD) and chlorhexidine gluconate (CHG). One or more inactive ingredients may comprise carriers, preservatives, emulsifiers, skin emollients and soothers and one or more other constituents.

The invention discloses a biological environmental antiviral cold prevention toothpaste and a preparation method thereof. The toothpaste contains the following components of: alkyl polyglucoside (APG), precipitated silica, lysozyme, neutral protease, hydrolyzed protein, immunoglobulin, recombinant human epidermal growth factor, strontium chloride, sorbitol, butylparaben, CMC (Carboxyl methyl Cellulose), xanthan gum, glucolase, enzyme, and a skin penetrating agent, etc. The invention is directed at the pathogenic mechanism that illness finds its way in by the mouth and nose, and the biologicalenvironmental antiviral cold prevention toothpaste with bio-environmental protection, virus resisting, cold prevention, as well as disease prevention and treatment effects is designed and prepared. During use, the toothpaste of the invention has cool, refreshing, icy and fragrant mouthfeel, and decomposes rapidly after entering the mouth. After tooth brushing, the ice-cold and refreshing feel remains for a long time, the bacteriostatic control time is long, the mouthfeel is refreshing, the taste is fragrant and bittersweet, and the spirit can be made cheerful, so that the toothpaste of the invention is beneficial to human health.

The invention discloses a medicine composition for promoting wound healing after operation and a preparation method thereof. The medicine composition is prepared from the following raw materials in parts by weight: 22-28 parts of natural indigo, 12-21 parts of radix notoginseng, 10-14 parts of Mongolian dandelion herb, 5-7 parts of Grosvenor momordica, 3-5 parts of liquorice root, 0.00005 part of recombinant human epidermal growth factor, 0.001 part of mannitol and 0.0005 part of human albumin. The medicine composition has obvious effects when being used clinically.

The invention provides a serum-free culture medium for mesenchymal stem cells. The serum-free culture medium specifically comprises a lipid mixture, glutamine, hydrocortisone, zinc sulfate, sodium butyrate, ferric citrate, vitamin C, recombinant epidermal growth factors, recombinant alkaline fibroblast growth factors, RGD oligopeptides, palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, lithium chloride, L-glutathione, a soybean pancreatin inhibitor, beta-mercaptoethanol, ethanolamine, sodium selenite, Hepes, sodium bicarbonate and a basic culture medium. The serum-free culture medium is free of human and animal resources, clear in chemical component, rich in nutrition, capable of achieving UC-MSCs rapid proliferation, and capable of solving the problem of cells number insufficiency and maintaining the biological features and immunophenotyping stability of the mesenchymal stem cells.

The invention discloses an endothelium culture system. The endothelium culture system comprises an endothelium culture medium and an endothelium additive, wherein the endothelium additive comprises components by concentration as follows: 5 ng/ml of a recombinant human epidermal growth factor, 10 ng/ml of hydrocortisone, 10 ng/ml of a recombinant human vascular endothelial growth factor, 5 ng/ml of a recombinant human basic fibroblast growth factor, 5 ng/ml of an R3 insulin-like growth factor, 50 mu g/ml of vitamin C, 50 mu g/ml of heparin, 10 mu g/ml of recombinant human insulin and 5 mu g/ml of human total iron transferrin. The endothelium culture system can promote growth activity of great vascular endothelial cells in a 5% serous environment, partially inhibit activity of fibroblasts and smooth muscle cells and reduce parenchyma cell pollution.