138results about How to "Does not affect biological activity" patented technology

The invention relates to the field of medicine preparation, in particular to a melittin lipidosome preparation. The lipidosome preparation is composed of one part of melittin, 5-40 parts of phospholipid, 1.3-10 parts of cholesterin and 10-140 parts of poloxamer. The invention also discloses the preparing method of the melittin lipidosome preparation. The melittin lipidosome preparation not only can delay the medicine release in the lipidosome, prong the circulating time in the blood and improve the bioavailability of the medicine, but also can obviously reduce the side effect of the medicine and improve the adaptability of a patient.

The invention discloses a long-acting recombinant human follicle-stimulating hormone-Fc fusion protein (hFSH-Fc) and a preparation method thereof. The hFSH-Fc is a dimerization fusion protein. An amino acid sequence of the hFSH-Fc comprises an hFSHbeta subunit, CTP (carboxy-terminal peptide), an hFSHalpha subunit, a flexible peptide linker and a human IgG (immunoglobulin G) Fc variant from the N terminal to the C terminal in sequence. The hFSH-Fc has longer half-life in vivo and smaller side effects than existing hFSH. The invention also relates to an application of a recombinant hFSH-Fc composition to preparation of drugs for treating and/or preventing infertility.

The invention provides a magnetic molecularly imprinted nano-particle as well as a preparation method and an application thereof. The nano-particle can perform specific recognition, trapping, separation and activity inhibition on target protein in a solution in vitro, more importantly, the nano-particle can enter a living cell rapidly under the action of a magnetic field and perform in-situ combination and activity inhibition on the target protein in the living cell, distribution of the nano-particles in the cell can be traced through a fluorescence microscope after fluorescence labeling, and quantitation can be performed through detection of fluorescence intensity.

The invention provides a bovine embryo vitrification tube swinging thawing and direct transplanting method and belongs to the technical field of embryo low-temperature biology. The method includes (1) freezing a bovine embryo and using a fine tube to first add a thawing liquid followed by air; (2) loading a freezing liquid followed by air; (3) loading a freezing liquid, placing the bovine embryo in the freezing liquid, and then loading air; (4) loading a thawing liquid, closing a tube opening, and performing freezing; (5) performing tube swinging thawing and direct transplanting, wherein the freezing liquid is an mPBS solution containing ethylene glycol (EG), polysucrose and sucrose, and the thawing liquid is an mPBS solution containing fetal bovine serum (FBS). According to the bovine embryo vitrification tube swinging thawing and direct transplanting method, the operation is simple and convenient, requirements for skills of operators are not high, the frozen bovine embryo can be directly transplanted after tube swinging thawing, the thawed embryo has a survival rate of 100% and an expansion rate of 94%, a pregnancy rate reaches 57% after transplantation, and the method is efficient, simple, easy to operate, and applicable to industrialized popularization.

A biotechnological method for efficient expression of interleukin-12 includes such steps as inserting the cDNA of IL-12's P40 subunit coding region into pcDNA3 eukaryon expression vector and the cDNA of IL-12's subunit coding region into pEE14 eukaryon expression vector to respectively configure the eukaryon cell's recombinant plasmids pcDNA3/p40 and pEE14/p35, cotransfecting host cells, adding G418 and MSX to screen culture medium, picking survival positive clone, and pressure amplification to obtain high-expression engineered cell strain IL-12.

The invention belongs to the technical field of biochemical engineering of antibiotics and surfactants, and in particular relates to a method for separating and extracting iturin A (Iturin A), which is produced by microbial fermentation and has surfactant properties and antibacterial activity. The fermented liquid is first treated with flocculants such as aluminum sulfate, polyaluminum chloride, polyferric sulfate, alum, and chitosan, and then iturin A is concentrated and extracted from the fermented liquid. The invention has the advantages of simple process route and low cost, and greatly improves the possibility of industrial and agricultural popularization and application of Iturin A.

The invention provides a separation method for silk fibroin peptides with small molecular weights. The separation method comprises the steps: degumming and dissolving silkworm silk to obtain a solution; adding the solution into a glucan G10 column, carrying out elution by using sterile water, and carrying out collection to obtain a desalted silk fibroin protein aqueous solution; freeze-drying thedesalted silk fibroin protein aqueous solution to obtain freeze-dried silk fibroin proteins; adding a freeze-dried silk fibroin protein solution into a glucan G50 column, eluting the silk fibroin proteins by using the sterile water, and collecting the silk fibroin proteins by using different tubes; screening the silk fibroin peptides of which the molecular weights are smaller than 1500Da in collecting tubes in the step (d), adding the silk fibroin peptides into a glucan G15 column again, eluding the silk fibroin peptides by using the sterile water, collecting the silk fibroin peptides by usingdifferent collecting tubes, and carrying out separation to obtain multilevel silk fibroin peptides with small molecular weights being 1500Da or smaller. The multilevel silk fibroin peptides with small molecular weights being 1500Da or smaller are obtained through dissolution with neutral salts and gradient separation of different molecular sieve columns. Meanwhile, the hydrophilicity of the silkfibroin peptides is made better by using a specific freeze-drying-dissolving method.

The invention provides a method for covalence coupling protein molecules onto surfaces of amino ceramic beads. The method is to use various activators to carry out activation for the amino ceramic beads and the protein molecules respectively, both of which are subsequently mixed to immediately accomplish the high-efficient covalence coupling of protein molecules onto the surfaces of the ceramic beads. Because of the different activators held by the activated protein molecules and the activated ceramic beads, no reaction occurs between the activated protein molecule and the activated ceramic beads, thereby efficiently avoiding the accumulation among protein molecules or the accumulation among the ceramic beads. In addition, the rapid and high-efficient reaction between mercapto group produced from activation and the activated maleimide group ensures the high coupling rate and good repeatability. The coupled ceramic bead can be widely applied to the fields of immune test, and cell selection.

The invention discloses a [<18>F] AlF marked positron emission tomography (PET) polypeptide probe and a preparation method thereof. The probe comprises TMTP1 polypeptide and NOTA, which are connected together, and the structural formulae of the TMTP1 polypeptide and the NOTA are shown in the description. According to the [<18>F] AlF marked positron emission tomography (PET) polypeptide probe and the preparation method thereof, a glycine is added to the TMTP1, and a G-TMTP1 polypeptide is designed, so that the polypeptide does not influence the biological activity after being radioactively marked; furthermore, 18F is taken as a radionuclide, and the G-TMTP1 polypeptide is marked by utilizing a [<18>F] AlF-NOTA method, so that the obtained [<18>F] AlF-marked PET polypeptide probe has excellent pharmacokinetics properties, the background cleaning rate is high, the liver uptake rate is low, in-vivo stability is good, the uptake rate of tumor site is high, highly metastatic tumor can be diagnosed specifically, and the polypeptide probe can be applied to diagnosis or therapeutic evaluation of highly metastatic malignant tumor.

The invention discloses a recombinant human chorionic gonadotrophin-Fc fusion protein (HCG-Fc for short) and a preparation method thereof. The HCG-Fc protein is a dimerized fusion protein, and the amino acid sequence of the fusion protein sequentially comprises an HCG beta subunit, an HCG alpha subunit, a flexible peptide joint and a human IgG Fc mutant from an N end to a C end. Compared with the existing chorionic gonadotrophin, the fusion protein has longer in-vivo half life, higher bioavailability and lower side effect. The invention also relates to an application of a recombinant HCG-Fc fusion protein composition in preparation of medicaments for treating and/or preventing infertility.

This invention relates to a medicine with rare earth transferring complex compound as medicine carrier. The invention provides a medicine comprising effective composition and carrier, wherein carrier is rare earth transferring complex compound. The medicine with rare earth transferring complex compound as carrier can be used as anticancer drug with orient transportation, for oral administration, and absorbent through lung permeation.

The invention provides a method for separating exosomes by a low-speed ultrafiltration centrifugation conjugated polymer precipitation technique. The method comprises the steps of performing concentration through a low-speed centrifugation and ultrafiltration centrifugal tube, and through combination with a polymer precipitation technique, separating the exosomes, wherein only a tabletop centrifuge common in laboratory is needed, and only the ultrafiltration centrifugal tube low in cost is needed. The operation method is simple and easy to operate, and the bioactivity of the exosomes is not influenced. The method also effectively solves the problems that compared with other cells, nerve cells are low in the quantity of secretion exosomes and difficult to separate and enrich. Besides, liquid containing the exosomes is effectively concentrated, and a polymer reagent which is used is greatly reduced. Compared with the prior art, the separation purity and the efficiency are higher, the maneuverability is higher, the content of the separated exosomes can be reduced, and the method has a favorable scientific application promotion value and favorable application prospects.

The invention discloses high-solubility compound protein powder and a preparation method thereof. Soybean protein isolate, whey protein concentrate and phospholipid are taken as main raw materials, stevia rebaudiana is processed into a stevia rebaudiana extract at low temperature, the effective ingredients and the bioactive ingredients of the stevia rebaudiana can be maximumly retained, the extraction rate is high, the flavor is pure, the color and cluster are natural, no licorice taste exists, the stevia rebaudiana extract is scientifically compounded with main materials, antifreeze polypeptides, dietary fiber flour, bee pupa hydrolysates and wheat malt powder are added, and then the compound protein powder which is appropriate in mouthfeel, good in solubility, comprehensive in nutrition, high in bioactivity and wide in suitable crowd is prepared. According to the preparation method, a non-thermal processing technique is adopted, the working efficiency is high, energy consumption is little, environmental friendliness is achieved, no pollution is generated, introduction of food additives and chemical auxiliaries is reduced, the food safety is effectively improved, the product quality is improved, the production cost is reduced, the low-carbon production objective is achieved, and large-scale and industrialized development can be achieved.

The invention discloses a method for extracting polypeptide by enzymolysis on oyster at low temperature, comprising the following steps of: carrying out enzymolysis on an edible part of a fresh oyster as a raw material with trypsase at the low temperature; extracting filter residue at low temperature again; combining filtrate of the two steps; and vacuum condensing to obtain a condensed solution containing the polypeptide. The method solves the problem that the enzymolysis of protein at high temperature is easy to inactivate a product of low-molecular weight peptide and broadens the application range of the oyster in the health-care foods.

The invention discloses a gene vaccine vector, and a preparation method and an application thereof; the gene vaccine vector is a supramolecular hydrogel formed through phosphatase catalysis of a small-molecular peptide; the small-molecular peptide has the structural formula represented by the formula (I), wherein when m=1, n=0, 1, 2 or 3; when n=1, m=1, 2 or 3. The gene vaccine vector after being loaded with DNA has the advantages of strong immunogenicity, large load capacity, no obvious toxicity and injectable immunity. The gene vaccine vector is mild in preparation conditions and simple in process.

The invention discloses a collagen aggregate based flocculent antibacterial hemostatic material and a preparation method thereof. The preparation method is characterized by comprising steps as follows: DCMC (dialdehyde carboxymethyl cellulose) and silver nitrate are blended, a mixed solution containing nano-silver is prepared through redox reaction, then certain amount of collagen aggregate and certain amount of polyvinylpyrrolidone are dissolved in the mixed solution sequentially, the mixture is subjected to processes including gradient dilution, dealcoholization, vacuum drying, impregnation, freeze-drying, moderate grinding, sterilization and the like, and the collagen aggregate based flocculent antibacterial hemostatic material is obtained finally. The material adopts the collagen aggregate more similar to a natural tissue collagen structure, and compared with common collagen materials, the prepared flocculent hemostatic material has excellent physical and chemical properties and biological properties such as structural stability, mechanical property, biological degradability, biocompatibility and the like, can promote wound healing and repair, has broad-spectrum antibacterial/antimicrobial activity of the nano-silver, has large contact surface with the wound surface, is high in hydrophilia and stickiness and can be used for hemostasis repair of body surface wound and general civil hemostasis emergency.

The invention provides a virus inactivation method for solution of globulin. An optimum proportion with a practical value of various parameters during virus inactivation with sodium caprylate is provided by a large number of researches on various conditions such as a temperature, a pH value, concentration of caprylate, concentration of protein and hatch time during the virus inactivation. Experiments prove that the virus inactivation method for the solution of the globulin has the advantages of simpleness, safety, rapidness, effectiveness, high protein recovery and no effect on the bioactivity of a target protein.

The invention discloses a method to manufacture biology polypeptide, especially a method to make natural human thymosin by gene series express method. It contains compounding two polynucleotide section, compounding double enzyme cut, T alpha 1 gene three times connecting in series, constructing high efficiency expression, constructing engineering fungus, expressing T alpha 1 polypeptide six series bodies in engineering fungus, purifying and cracking. The invention could abundantly express aim albumen, and it has simple technology, easy to operate and low cost.

The invention discloses a method for preparing human LIGHT-Fc fusion protein by using a pichia expression system, comprising the following steps of: (1) constructing recombinant expression plasmid pPIC9K-LIGHT-Fc; (2) preparing and screening expression human LIGHT-Fc fusion protein yeast engineering bacteria; and (3) expressing and evaluating the human LIGHT-Fc fusion protein in pichia. The invention makes up the deficiency of the traditional human LIGHT preparation method and the expression mode of a prokaryotic expression system, otherwise, if being used for producing industrial human LIGHT protein, the invention has the advantages of simple operation, short growth cycle of raw organisms, large production scale (capable of carrying out high-density fermentation), low extraction cost, high activity of products, and the like and has important industrial application prospect and actual significance.

The invention discloses a method for preparing TiO2@Au core-shell structure in a template method. The method comprises the following steps: preparing PS@Au in a template method, coating the PS@Au witha layer of amorphous titanium dioxide to obtain TiO2@PS@Au, and calcining the TiO2@PS@Au to obtain TiO2@Au. PS is used as a template, Au is wrapped in the TiO2, a uniform and stable TiO2@Au core-shell structure can be obtained after the calcinations, and the dispersion property is good. The core-shell structure is a hollow structure, the weight is light, and the surface area is relatively large under the same mass. Nano gold can be combined with various biological macromolecules, the bioactivity is not influenced, the nano gold can be applied to the biological catalysis and biological sensing, the TiO2@Au obtained by the invention can be used as an enzyme reactor, compared with the method only adopting the pure titanium dioxide as the carrier, the separation of the nano gold in the subsequent reaction can be avoided, the advantages of the titanium dioxide semiconductor can be played, and the overall catalytic efficiency can be increased.

The present invention relates to nano filtering technology of desalting and concentrating oligopeptide solution, and belongs to the technology of purifying and concentrating protein and peptide. Milkprotein, silk protein, soybean protein, cereal protein, egg protein, meat protein, fish protein and cotton seed protein material is treated to obtain protein solution; the protein solution is temperature and pH regulated, hydrolyzed, via enzyme process or acid process to reach required degree of hydrolysis; and through centrifugation and filtering, oligopeptide solution is obtained. The oligopeptide solution is desalted and concentrated and the concentrated solution is spray dried to obtain the oligopeptide powder product soybean peptide, corn peptide, silk peptide, etc.

The invention belongs to the technical field of immune cell culture, and particularly relates to a cell culture solution and an NK cell culture method. The provided cell culture solution includes various kinds of culture solutions including an ALyS505NK-AC culture solution and an ALyS505NK-EX culture solution, wherein the ALyS505NK-AC culture solution containing IL-2 and IL-5 and the ALyS505NK-EX culture solution includes IL-2. The invention further provides the NK cell culture method, the biological activity of NK cells obtained by means of the method is high, the number of amplification is large, therefore, the provided culture method is simple in operation, the survival rate of the NK cells is high, the number of amplification is large, and the culture solution and the culture method can meet various application requirements of clinical or academic research.

The invention discloses a method for extracting a paracrine factor from adipose-derived stem cells. The method comprises the following steps: taking a collected fat sample, and washing the fat samplewith normal saline for 1-3 times; adding a proper amount of adipose cell digestive juice for digestion; performing centrifugation, resuspending cell precipitate, and performing primary culture with aserum-free medium; performing subculture; performing enlarged culture; and performing extraction with a paracrine factor extracting solution, and performing purification to obtain the paracrine factor, wherein the paracrine factor extracting solution is prepared from a PBS buffer solution, L-glutamine, D-glucose and L-ascorbic acid, and the concentrations of the L-glutamine, the D-glucose and theL-ascorbic acid are 1-3mmol/L, 10-20 micromoles/L and 10-100 micromoles/L respectively. The method disclosed by the invention can stably extract the adipose-derived stem cells from adipose tissues, and extract the paracrine factor of the adipose-derived stem cells by utilizing the adipose-derived stem cells, so that the cell quality and the factor extraction efficiency are stable; and the adipose-derived stem cells are separated and cultured for a long time in a GMP environment, do not contact any animal-derived component, and can efficiently express adipose-derived stem cell markers such as CD29, CD44 and CD105.

The invention discloses a thrombus filter capable of realizing delayed recovery and a preparation method of the thrombus filter and particularly discloses a blood compatible coating. The blood compatible coating comprises a polyacrylic acid layer and a polyethylene glycol layer, and 2-methacryloyloxyethyl phosphorylcholine is covalently linked with the surface of the polyethylene glycol layer. Theinvention further discloses medical apparatus such as the thrombus filter coated with the blood compatible coating. According to the technical scheme, thrombogenesis and endothelial tissue growth canbe better inhibited. The whole surface coating is adjustable in thickness and surface grafting amount and can well bear stress and strain actions in apparatus assembly and release processes, molecules are firmly combined without falling off, and mechanical properties of the apparatus are not affected.