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

A process for preparing high-capacity microreticular agarose gel medium includes such steps as dispersing the granular calcium carbonate as hole-forming agent in the aqueous solution of agarose to obtain suspension, adding it to salad oil, adding emulsifier Span80, emulsifying while stirring, quick cooling for sphericizing, adding epoxy chloropropane, cross linking, reducing with sodium borohydride, and using diluted hydrochloric acid to remove granular calcium carbonate.

The present invention discloses a performance improved recombination staphylococcus aureus protein A affinity ligand and a construction method thereof. The present invention adopts a molecular biology method. A sequence B of a nature protein A is selected for molecular transformation. A C-terminal of the sequence B is added with two cysteines, so that the protein A can pass through double-locus coupled chromatography matrix to stabilize the connection. Six glycines are added to the end of a second Loop of the sequence B to increase the length and reduce the binding force with an antibody, so that elution conditions are mild. On this basis, resistance performance to high concentration base of the protein A is transformed. Asparagines and phenylalanine at 23rd and 30th positions of the sequence B are respectively replaced by threonine and alanine to obtain a sequence Z with higher alkaline resistance properties. Then, isocaudarner is used for connecting sequence Zs of different numbers head-to-tail in series. The efficient expression system of e. coli is used for overexpression. The expressed recombination protein A is coupled to agarose matrix preparation affinity chromatography fillers and is used for purifying antibodies. Results show that the recombination protein A affinity ligand prepared by the present invention is good in elution performance and alkali resistance.

The invention discloses a preparation method of a hydrophobic charge induction chromatography medium with large capacity, and belongs to the protein chromatography separation technology in the field of biochemical engineering. The chromatography medium refers to the end of a space arm that is coupled with imidazol compounds as a chromatography matrix. The preparation method is that: the medium is activated after dimethyl sulfoxide, epichlorohydrin and sodium hydroxide solution are sequentially put into agarose gel medium; the activated medium is coupled with the imidazol compounds matrix in the alkaline dimethyl sulfoxide solution; finally, the epoxy left on the medium surface gets the needed chromatography medium after being reduced. The adsorption capacity of the chromatography medium to the protein achieves a moist medium of 80mg/ml and keeps relatively stable within a broader ionic strength scope from 0.1 mol/L to 1.0 mol/L; at the same time, the protein elutropic model which is finished by depending on the regulation of PH between 4.0 and 6.0 is more moderate, thereby having the wide application prospect in the separation and purification of biomacromolecule such as the protein, etc.

The invention relates to a method for preparing composite TiO2 copolymer microspheres and application of the composite TiO2 copolymer microspheres in natural product separation and purification. According to the method, an oil phase containing surface-modified TiO2, benzoyl peroxide serving as an initiator, GMA serving as a monomer, DVB serving as a crosslinking agent and the oil phase of a pore forming agent is added into a composite aqueous phase containing gelatin and sodium chloride, and the mixture is heated to cure to form microspheres; and beta-CD is directly immobilized on the microspheres serving as a immobilization matrix in the presence of sodium hydride to form the composite TiO2 copolymer microspheres. The copolymer microspheres can be used as an expanded bed adsorption medium to separate isoflavones from soybean waste molasses and purify the isoflavones. The method has the advantages of simple preparation process and easy control and amplification. The prepared TiO2 copolymer microspheres have the advantages of good medium sphericity, mild elution conditions, high biocompatibility, excellent hydrophilicity and wide application prospect in the natural product separation and purification.

The invention provides a high-efficiency specificity enriched hemagglutinin peptide HA mark recombinant protein immunoaffinity purification enriching column as well as a preparation method and an application thereof. The immunoaffinity purification enriching column provided by the invention comprises an activated agarose filler coupled with HA specificity polyclonal antibody which is subjected to immunoaffinity purification and a plastic column carrying the immunoaffinity filler. The immunoaffinity purification filler coupled HA specificity polyclonal antibody is extracted by using an immunoaffinity method; and the immunoaffinity purification enrichment column is obtained by carrying the immunoaffinity filler into a specific plastic column. The immunoaffinity purification enrichment column enriched HA mark recombinant protein is high in efficiency, strong in specificity and temperate in elution condition, can be repeatedly utilized and is an ideal material for separating, purifying and enriching the HA mark recombinant protein at present.

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 discloses an application of modified resin in the process of boron removal. The structure of the modified resin is shown by a formula in the specification, wherein R represents macroporous Merrifield resin with the model number of G20M1254. An experiment proves that the modified resin is relatively good in boron removal capacity in an aqueous solution, and the elution condition is mild, so that the service life of the resin is greatly prolonged. Moreover, the application disclosed by the invention is simple and convenient to operate and easy to implement, is low in equipment requirement, can greatly save the cost and has a practical application value. A hydroxyl radical and a benzene ring of a phenolic compound in the structure of the modified resin are positioned on the same plane, so that o-diphenol can be complexed with boric acid smoothly. The boron removal capacity of the modified resin can reach 6.16mg/g, and the environmental pollution caused by the modified resin can be reduced.

The invention provides a D-tag tagged recombinant protein immunoaffinity purification and enrichment column capable of efficiently and specifically enriching flag peptide tagged protein and a preparation method and application thereof. The provided immunoaffinity purification and enrichment column comprises an activated agarose filler coupled with an immunoaffinity-purified D-tag specific polyclonal antibody and a plastic column loaded with the immunoaffinity filler, wherein the D-tag specific polyclonal antibody coupled with the immunoaffinity filler is extracted by an immunoaffinity method. The immunoaffinity purification and enrichment column is prepared by loading the immunoaffinity filler into the special plastic column; and the immunoaffinity purification and enrichment column is high in efficiency and strong in specificity while enriching D-tag tagged recombinant protein, is mild in diluting conditions, can be repeatedly utilized and is an ideal material for separating, purifying and enriching the D-tag tagged recombinant protein.

The invention relates to a post-extraction technology of a chicken alpha-disruptor, and belongs to the technical field of production of medicines for livestock. The technology comprises the steps of S1, performing centrifugation on fermentation liquid containing the chicken alpha-disruptor, and reserving supernatant; S2, filtering the supernatant with a 50KDa ceramic membrane, filtering filtrate through a 10KDa ceramic membrane, and collecting trapped liquid; S3, adding a Cu<2+>-IDA-glucan carrier and a NaCl solution, with the same volume to the trapped liquid in the step S2, and performing anadsorption reaction to obtain reaction liquid; S4, adding a phosphate buffer solution with the same volume to the reaction liquid in the step S3, performing uniform mixing, performing filtering withthe 50KDa ceramic membrane, and collecting trapped liquid; S5, adding eluent to the trapped liquid in the step S4, then performing filtering through the 30KDa ceramic membrane, and collecting filtrate; and S6, filtering the filtrate in the step S5 through the 10KDa ceramic membrane for desalinating, and collecting trapped liquid to obtain the chicken alpha-disruptor. The yield is 61% or above, thepurity is 98% or above, and the activity yield is 96.73% or above.