34results about How to "High recovery rate of activity" patented technology

The invention discloses a method for preparing a freeze-dried human blood coagulation factor VIII. The method comprises the following process of: dissolving by taking water for injection, comprising 3-10 IU (International Unit)/ml of heparin, as a heparin sodium solution and cryoprecipitating; performing PEG (Polyethylene Glycol) precipitation and taking supernatant; performing centrifugal filtration; performing S/D (Solvent/Detergent) inactivation at the temperature of 24-26 DEG C; performing DEAE (Diethylaminoethyl) Sepharose Fast Flow chromatographic column balance, adsorption, washing andelution; performing molecular membrane ultrafiltration; preparing, removing bacteria, sub-packaging, freeze-drying and capping; and dry-heating at the temperature of 99.5-100.5 DEG C and inactivating. In the invention, the process method of combining the PEG precipitation and an ion exchange chromatography technology is adopted; the method is easy and convenient to operate; the F VIII active recovery rate is increased; miscellaneous proteins can be removed on a large scale; the product yield reaches over 60 percent; and the specific activity of the product reaches 5 IU/mg and is obviously greater than a value which is not less than 1 IU/mg stipulated in the pharmacopeia. Meanwhile, the PEG residue is 0.08g/L which is obviously less than the value which is less than or equal to 0.5 IU/mg stipulated in the pharmacopeia, so that Al<3+> residues in the final preparation of an Al(OH)3 gel method are avoided; the product has high purity and high safety; and the quality of the final product is obviously improved.

This invention discloses a method for manufacturing silk fibroin nanoparticles fixed with enzyme from silk fibroin, which comprises the steps of: (1) completely mixing the enzyme and regenerated silk fibroin solution; (2) injecting into a water-soluble organic solvent under high speed stirring to obtain white crystalline silk fibroin nanoparticles fixed with the enzyme; and (3) centrifuging or filtrating to remove the organic solvent to obtain crystalline silk fibroin nanoparticles fixed with the enzyme. The average particle size of the nanoparticles is 35-125 nm, and the activity recovery is as high as 70%. The fixed enzyme has a high thermal stability, and is not easy to be decomposed by proteinase, thus can largely reduce or even eliminate the immunogenicity of the enzyme. The nanoparticle fixed with the enzyme has important applications in sustained drug release, industrial enzyme reactor, food additives and cosmetics.

The invention discloses a method for preparing reusable magnetic nanometer immobilized enzyme. The method comprises the following steps: nanometer Fe3O4 powder with surface on for amino group adsorption is prepared with a coprecipitation method; by using the surface groups, the nanometer Fe3O4 powder reacts with enzyme molecule amino group under the action of cross linking agent, and accomplishes cross linking process, then solid-liquid separation is performed under the action of external magnetic field, immobilized enzyme is recycled, and the magnetic nanometer immobilized enzyme which can be reused can be obtained. The magnetic nanometer immobilized enzyme of the invention has the advantages of high protein load capacity, high enzyme activity, convenient recycling and stable chemical property, and is suitable for the catalytic field, the degeneration of environment pollutant and the synthesizing process of energy material.

The invention discloses a dry heat treatment method for a human coagulation factor VIII preparation. Before a freeze-drying step, the stabilizer is added into human coagulation factor VIII solution, wherein the stabilizer comprises human serum albumin, Ca2+ soluble salt, amino acid, sodium citrate and sodium chloride; and the concentration percentage of the human serum albumin is 0.5 to 5 percent and the concentration of Ca2+ is 1mmol/L. The invention also discloses the dry heat treatment stabilizer for the human coagulation factor VIII preparation. The dry heat treatment method is performed for 72 hours at 80 DEG C and is effective on both lipid envelop virus and non-lipid envelop virus. Usually, heat treatment is the last step. The method is low in cost and higher in maneuverability and controllability.

The invention relates to a biomimetic affinity purification method of a para-aminophenamidine biomimetic affinity ligand. The biomimetic affinity purification method of the para-aminophenamidine biomimetic affinity ligand comprises preparation of a biomimetic affinity material as well as biomimetic affinity purification on metalloprotease (MP) by a para-aminophenamidine agarose gel column. The purification method is simple in flow, short in cycle and high in efficiency, and the purity is up to 98.7 percent.

The invention discloses a preparation method for magnetic sodium alginate-immobilized laccase, and belongs to the technical field of immobilized enzyme preparation. According to the method, a laccase enzyme liquid is prepared by fermenting a laccase production strain, magnetic particles are mixed to immobilize the laccase enzyme liquid in combination of sodium alginate in an auxiliary manner, and drying is carried out after oscillation adsorption, so as to obtain the magnetic sodium alginate-immobilized laccase. The method is simple in process and low in cost, the activity of the prepared magnetic sodium alginate-immobilized laccase is 250 IU/mL or above, the total output is increased to 85-90%, and both the activity and the total output reach a higher level; moreover, through mixing of the nanoscale magnetic particles, the adsorption is more easy to realize, and separation is easy, so that the recovery rate of the immobilized laccase is improved, the stability is remarkably enhanced, and the laccase has excellent performance.

The present invention discloses a preparation method of nano magnetic chitosan immobilized enzyme. Said method includes the following steps: making nano magnetic chitosan particles whose surface contains acylchloro functional group be dispersed in enzyme-bearing solution, stirring them to make reaction for 2-8 hr at 20-25 deg.C, making solid-liquid separation in magnetic field, using phosphoric acid buffer solution to wash the magnetic precipitate for three times so as to obtain the required nano magnetic chitosan immobilized enzyme, the concentration of enzyme in enzyme-bearing solution is 0.01-0.1%. and the solid-liquid ratio of nano magnetic chitosan whose surface contains acylchloro functional group and enzyme-bearing solution is 1:50-1:200 g/ml.

The invention discloses a separation and purification method of recombinant human interferon beta 1a. The separation and purification method comprises the following steps: 1) carrying out blue gel chromatography on a sample, collecting active peaks of the recombinant human interferon beta 1a; 2) carrying out ultrafiltration and desalination on a product obtained by the blue gel chromatography; 3)carrying out Q-Sepharose Fast Flow anion exchange chromatography on the ultrafiltration and desalination product, balancing a column by using 20mmol//L Tris-HCl buffer liquid with the pH of 8.3-8.7, after sampling, eluting with the buffer liquid to a base line, and then eluting by using 20mmol/L NaCl-containing Tris-HCl buffer liquid with the pH of 8.3-8.7, and collecting the active peaks of the recombinant human interferon beta 1a; and 4) carrying out S-200 molecular sieve chromatography on the product obtained by the anion exchange chromatography so as to obtain the recombinant human interferon beta 1a. By using the purification method, a target protein has small possibility of inactivation, and the purity and biology specific activity of the target protein obtained by purification are high.

The invention discloses a process for preparing a human prothrombin complex concentrate by adopting a flow adsorption method. The process is characterized in that a plurality of cylindrical containerswith stirrers are connected in series to form a natural flowing way; gels which are balanced in advance are placed in the various containers; a certain batch of to-be-processed blood plasma is storedabove the first container; the gels are injected quantitatively firstly in the first container, then the gels are stirred and adsorbed for a unit time, and the adsorbed blood plasma is discharged into the second container at a certain flow speed; simultaneously, other to-be-processed blood plasma flows into a first adsorption tank at the same flow speed, so that dynamic balance of the blood plasma quantity in the first container is kept, and at this time, adsorption and separation of the blood plasma in the first container are carried out simultaneously; by analogy, after the all blood plasmapasses through, outlets of the various containers are closed, and washing and elution on the gels are carried out. The blood plasma quantity is not limited by the volume of the containers, the process is easily controlled, an FIX activety recycling ratio is higher, and the risks of leakage of the gels and crossed pollution are avoided.

The invention relates to the field of enzyme immobilization, and in particular relates to an immobilized cutinase and a preparation method and an application thereof in removal of phthalic acid esters in water. The immobilized cutinase is fixed on a porous gold nanomaterial through a physical absorption function by taking the porous gold nanomaterial as a carrier, wherein the adsorption quantity of the cutinase on the porous gold nanomaterial is more than 30mg/g; the bore diameter of the porous gold nanomaterial is between 40nm and 50nm. The preparation method comprises the following steps: adding the porous gold nanomaterial into a cutinase solution; adsorbing in an oscillation manner at 45 DEG C to 55 DEG C; and performing washing, freezing and vacuum drying, thereby obtaining the immobilized cutinase. The immobilized cutinase can be used for degrading the phthalic acid esters in the water, and is good in biocompatibility, high in adsorption quantity and good in adsorption stability.

The invention relates to a phospholipid nano-disk chromatography medium and a preparation method and application thereof. The phospholipid nano-disk chromatography medium comprises a charged phospholipid nano disk and an affinity matrix; the charged phospholipid nano disk comprises phospholipid and a film scaffold protein with an affinity label; the charged phospholipid nano disk is connected withan affinity matrix through the affinity label. The phospholipid nano-disk chromatography medium has both binding capacity and protection capacity to film proteins, so that the actual system containing the film proteins can be purified and reconstructed on a chromatography column in one step; and the phospholipid nano-disk chromatography medium has reversibility and regeneration performance; the phospholipid nano-disk chromatography medium solves the problems of serious protein activity loss, long and complicated steps and time-consuming and labor-consuming operation in the traditional film protein research technology, is simple and efficient to operate, improves the film protein activity recovery rate by more than 20 times, and shortens the treatment time by more than 95%.

The invention is applicable to the technical field of biochemistry, in particular to a calibrator for a biochemical calibrator and the biochemical calibrator. Each liter of the matrix for the biochemical calibrator comprises the following components of 10 to 100 mmol of a pH regulator, 2-50 g of a saccharide protective agent, 2-100 g of a polymer protective agent, 5-50 g of a protein protective agent, 0.5 to 5 grams of surfactant, 5 to 20 millimoles of amino acid, 0.02 to 50 millimoles of antioxidant, 0.05 to 2 millimoles of protease inhibitor, 5 to 30 grams of alcohol organic solvent, 0.2 to0.5 gram of preservative, 8.5 to 9.5 grams of sodium chloride and 0.1 to 5 millimoles of magnesium salt and/or zinc salt. According to the calibrator for the biochemical calibrator, when the analysiscomponents are added into the matrix for the biochemical calibrator, the components in the matrix for the biochemical calibrator can play a better role in protecting the analysis components, particularly, the structural variation caused by water loss of the analysis components in the freeze-drying process can be avoided, and the effect of the analysis components is effectively reserved.

The invention relates to an efficient preparation method of a papain preparation. The method comprises the following specific operation steps: S1, preparing a papain preparation crude product; S2, preparing a papain crude enzyme product solution and a papaya lipase crude enzyme product; S3, preparing a papaya lipase preparation; and S4, isolating and purifying papain. According to the efficient preparation method of the papain preparation disclosed by the embodiment of the invention, the papain and the papaya lipase can be obtained simultaneously by isolation and purification of papaya primary pulp, complete utilization of resources is realized, the preparation steps are simple, the design is reasonable, the operation is simple, the influence on the activity of the enzymes in the preparation process is relatively small, the prepared papain and papaya lipase are high in purity, high in activity and high in enzyme activity recovery rate; and reagents can be recycled after isolation is performed with a strong cation exchange method, so that the reagents are repeated used, the cost is effectively reduced, the preparation can be expanded, and the industrial requirements are met.