32results about How to "Mild elution conditions" patented technology

The invention discloses a biomimetic monolithic material with the affine selectivity similar to that of a protein A, and a preparation method thereof, wherein the material is capable of purifying, separating and immobilizing antibodies. N, N'-di-(2-aminoethyl) oxamide and 4-mercapto-phenylboronic acid are used as common functional monomers, tri-(2, 3- epoxy group propyl group) isocyanate is used as a monomer, polyethylene glycol is used as a porogenic agent, and reaction is carried out in a mode of in-situ ring-opening polymerization to prepare the biomimetic monolithic material with the affine selectivity similar to that of the protein A. The material can be combined with antibodies specifically, and can be applied to the fields of antibody purification, separation, immobilization and the like. The material has the advantages of high selectivity, low cost, good stability, easiness in elution, reusability and the like, and the immunity affinity and the selectivity of the antibodies are not affected.

The invention relates to a method for separating immune globulin IgG from human serum. The method comprises the following steps: 1) diluting the human serum by a buffer solution, adjusting a pH to 6.0to 8.0 and filtering by a filter membrane to obtain a human serum sample; 2) feeding the human serum sample into a chromatographic column, washing by a balancing buffer solution, then eluting by a eluting buffer solution and collecting eluent to obtain the immune globulin IgG solution, the chromatographic column is filled with a combined type bionic chromatography medium, the combined type bionicchromatography medium is prepared from a chromatography matrix and combined type ligand, the chromatography matrix is a hydrophilic porous microsphere with hydroxyl, and a sequence of the combined type ligand is phenylalanine-tyrosine-glutamine-5-aminobenzimidazole; 3) performing utlrafiltration and concentration on the immune globulin IgG solution to obtain the immune globulin IgG. The method can improve an efficiency and a purity for separating the immune globulin IgG from blood.

The invention discloses a method for adsorbing and separating human serum albumin by expanded bed adsorption based on a mixed mode, which can directly separate and recombine human serum albumin from a yeast fermentation solution, and belongs to the protein separation technology in the biochemical field. The method comprises the following steps: (1) preprocessing a fermentation solution; obtaining the yeast fermentation solution containing the human serum albumin, adding sodium caprylate, and heating and deactivating protease; (2) performing expanded bed adsorption, directly separating the yeast fermentation solution by using an expanded bed filled with a mixed-mode adsorption agent, and collecting elution peaks; and (3) desalting and drying: desalting a collected solution, freeze drying to obtain the human serum albumin with the purity greater than 95 percent. The invention is characterized by developing a novel separation process, so that the high-purity recombined human serum albumin can be directly separated from the yeast fermentation solution. A critical point of the invention lies in adopting the expanded bed adsorption agent based on the mixed mode, cells do not need to be removed from the yeast fermentation solution, the ion strength does not need to be adjusted, the elution condition is moderate, and the method has the characteristics of simple process, high separation efficiency and high yield.

The invention belongs to the protein chromatographic separation technology in the field of biochemical industry and discloses a mixed model chromatographic method for separating human serum albumin from yeast fermentation liquid. The method includes steps: 1) fermentation liquid pretreatment, to be more specific, removing yeast cells by centrifuging the yeast fermentation liquid which contains the human serum albumin, adding sodium caprylate, heating to remove impurity proteins and inactivating protease, performing centrifugal separation, and taking supernatant; 2) column chromatography, to be more specific, adopting a mixed mode medium with tryptophan as ligand, performing fixed bed chromatographic separation of the supernatant, and collecting elution peaks, wherein a sample loading pH value is 4.0-4.5, and an elution pH value is 7.0-9.0; 3) desalting and drying, desalting collected liquid, performing freeze drying to obtain the human serum albumin with purity larger than 95%. The mixed model chromatographic method is characterized in that by development of a novel separation process, the high-purity human serum albumin can be directly separated from the yeast fermentation liquid. By adoption of the mixed mode medium with tryptophan as the ligand, ion strength adjustment of the yeast fermentation liquid is avoided, adsorption under acid conditions and elution under neutral and weak alkali conditions are realized, and the method has advantages of mild elution conditions, simple process, high separation efficiency and high yield.

The invention discloses an affinity chromatography medium employing tetrapeptide as a functional ligand and a preparation method of the affinity chromatography medium. The method comprises the following steps: adding dry matrix and allyl bromide to a dimethyl sulfoxide solution, activating, and reacting activating matrix with N-bromo succinimide; enabling bromo alcoholized matrix to react with hexamethylendiamine to obtain amino activating matrix; sequentially washing with deionized water, absolute ethyl alcohol and anhydrous N,N-dimethyl formamide, adding an N,N-dimethyl formamide solution containing tetrapeptide, 2-(7-azobenzotriazole)-N,N,N',N'-te-tramethyluronium hexafluorophosphate and N,N-diisopropylethylamine, and coupling a tetrapeptide ligand; and putting a medium coupled to tetrapeptide in a mixed liquid of sodium acetate and acetic anhydride to obtain the affinity chromatography medium employing tetrapeptide as the functional ligand. According to the novel chromatography medium developed by the method, a functional group is tetrapeptide composed of tyrosine, phenylalanine, arginine and histidine, and is designed on the basis of a protein A binding site of an antibody Fc segment; the antibody binding selectivity is greatly improved; and the affinity chromatography medium can be applied to efficient separation of an antibody.

The invention discloses a preparation method of a microsphere PGDT separation medium with two types of pores. The method is to mix the monomer, cross-linking agent, liquid porogen, and initiator first, then add the solid porogen and mix evenly, and form microspheres through suspension polymerization, extract the microspheres with absolute ethanol, and pickle , and dry, two types of pore-type separation media can be obtained. It is characterized in that: the solid porogen is calcium carbonate, and the dosage accounts for 10-3% of the volume content of the reaction mixture; the volume ratio of the solid porogen to the liquid porogen is 0.5-1; the dosage of the porogen accounts for 3% of the volume content of the reaction mixture. 20-60%; the amount ratio of the two cross-linking agents is 0.5-2, and the ratio of the amount of cross-linking agent to the amount of monomer is 0.3-0.6; under normal pressure, the polymerization temperature is controlled at 65-85 between ℃. The separation medium prepared by the method can be used for the separation of biomacromolecules such as proteins after modification, has large adsorption capacity and mild elution conditions, and has good application prospects in large-scale preparative separation of biomacromolecules.