9 results about "Sepharose" patented technology

The invention relates to a pharmaceutical composition for reducing dentin hypersensitivity and a preparation method thereof, and belongs to the technical field of biological medicine, the pharmaceutical composition comprises chanterelle extract, rabdosia rubescens leaf extract, eggshell membrane protein peptide and bifidobacterium thermophile fermentation extract; the cantharellus cibarius extract is obtained by sequentially performing enzymolysis on cantharellus cibarius through compound enzyme, ginger protease and serratipeptidase and purifying enzymatic hydrolysate through an AB-8 macroporous resin column. The rabdosia rubescens leaf extract is obtained by ultrasonically extracting rabdosia rubescens leaves with an ethanol water solution and purifying an extracting solution with a composite filler adsorption column. The eggshell membrane protein peptide is obtained by carrying out enzymolysis on a shell membrane by virtue of ginger protease and ficin and purifying an enzymatic hydrolysate by virtue of a G-25 sephadex column. The fermentation extract of the bifidobacterium thermophilus is obtained by purifying fermentation liquor of the bifidobacterium thermophilus through an SP agarose gel FF column. All the components are compounded according to a specific ratio and have a synergistic effect, so that the dentin hypersensitivity can be relieved safely in a multi-effect manner.

The invention discloses a method for purifying atosiban. The method sequentially comprises the following steps: dissolving an atosiban crude product with an acetic acid aqueous solution, and adjusting the pH value to 4.0-5.5; carrying out primary purification by adopting SP Sepharose FF cation exchange resin, and removing a large amount of deleted peptide impurities; 0.5-1.0 mM of TCEP is added for reduction treatment, and disulfide bond mismatch is corrected; carrying out fine purification through C18 reversed-phase preparative chromatography, and removing oxidation by-products; and finally, carrying out nanofiltration desalination and freeze drying to obtain the high-purity atosiban. According to the method disclosed by the invention, the purity of the final product is more than or equal to 99.5%, the total yield is more than or equal to 65%, the maximum single impurity is less than or equal to 0.15%, the unit consumption of acetonitrile is less than or equal to 12 L / kg, the amplification verification of 100 mg to 500 g is completed, the process is stable, the cost is low, the industrial applicability is high, and a reliable technical scheme is provided for the large-scale production of the atosiban raw material medicine.

Anchor molecules for peptide synthesis in aqueous solutions are disclosed. Wherein the anchor molecules are independently present and may be covalently coupled to a variety of hydrophilic solid supports, such as CM agarose and carboxymethyl cellulose, by amido bonds under aqueous phase conditions using a coupling agent such as EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide) prior to the start of peptide synthesis. The anchor molecule has a general structure of Fmoc-AA-CH2-Ph-CH2-Ph-CH2-NH-(CH2) n-NH2, wherein Fmoc-AA represents an amino acid protected by Fmoc, and the general structure of the anchor molecule is shown as Fmoc-AA-NH-(CH2) n-NH2. The anchoring strategy includes an acid labile ester bond located between the carboxyl terminal of Fmoc-AA and the benzyl alcohol group of the solid support, which ester bond remains stable during peptide extension and is cleavable under strongly acidic conditions, achieving final peptide release. The anchor molecule may be functionalized to interact with the ion exchange resin. The present invention enables efficient solid phase peptide synthesis (SPPS) in an aqueous phase system, enhances compatibility with hydrophilic resins, and provides improved environmental sustainability by reducing dependence on organic solvents.

The application discloses a kind of white snail glycosaminoglycan-growth factor composite active substance directional extraction purification process, belong to biological active substance extraction technical field.Core breakthrough is in at: through-10 ℃ acetone circulation degreasing and 20kHz ultrasonic coordination papain-neutral protease compound system (mass ratio 1:0.8), realize glycosaminoglycan and growth factor synchronous efficient release, extraction rate reaches 97.2%;Using DEAE-Sepharose Fast Flow and Q Sepharose Fast Flow double column chromatography gradient elution, combined with 1000Da ultrafiltration membrane refining, composite active substance purity is ≥99.6%;Through-Vc mild depolymerization technology regulates glycosaminoglycan molecular weight to 3.8~23.8kDa, retains its heparin-like structure activity;Using porous medium enhanced vacuum freeze-drying process, so that composite active substance activity retention rate is ≥98.5%.Product glycosaminoglycan content is ≥35mg / g, growth factor (EGF class) activity is ≥1200IU / mg, DPPH free radical clearance rate is ≥65%, to skin fibroblast proliferation promotion rate is ≥72%, no organic solvent residue and heavy metal pollution.

The application provides a detection reagent for frozen semen of Huai pig, and first acquires the raw semen, and then carries out the following steps: ammonium sulfate fractionation precipitation, DEAE cellulose filter gel column chromatography, CM Sepharose FastFlow gel chromatography and Sephadex G-100 chromatography to separate and purify the NAGase of the semen of the Huai pig, and monoclonal antibodies JN21 and JN37 are prepared by taking the NAGase as an antigen, and the monoclonal antibodies can specifically recognize the NAGase of the Huai pig. The prepared double-antibody sandwich ELISA kit can rapidly and quantitatively detect the NAGase content in the semen of the Huai pig, and provides a new thought for Huai pig breeding and purification and rejuvenation. The kit is convenient to operate, low in use cost, good in repeatability and suitable for being widely applied.

This invention belongs to the field of biotechnology. Specifically, it discloses a method for preparing recombinant protein using rice seeds. The method involves dehulling rice seeds to obtain rice flour, which is then enzymatically hydrolyzed using cellulase and neutral protease in a buffer solution. This process efficiently hydrolyzes the cellulose in the rice cell wall and some impurities. The buffer solution reduces protein salting out and increases the dissolution rate of recombinant human lactoferrin. Activated carbon is used to initially adsorb and remove impurities such as natural pigments and phenolic substances from the rice seeds. The mixture is then subjected to chromatography using CM Sepharose FF, SP Sepharose XL, and CM Sepharose FF columns, effectively improving the purity and recovery rate of the recombinant human lactoferrin. Simultaneously, the enzymatic hydrolysis and fractionation chromatography effectively reduce the iron saturation in the recombinant human lactoferrin.

The invention relates to a method for separating plasma small extracellular vesicles (sEV) subsets and application of the method in treatment of sepsis cardiomyopathy, and belongs to the technical field of extraction of plasma small extracellular vesicles. According to the method disclosed by the invention, propylene sephadex S-400 and S-500 are combined to be filled into a chromatographic column, and different sEV subgroups in plasma are separated through a dual-size exclusion chromatography (d-SEC), so that the sEV with high purity, complete structure and heterogeneity is obtained. According to the method, more different high-purity plasma subgroups sEV can be conveniently, economically and practically obtained, and good technical support and method models are provided for clinical and basic research. Furthermore, it is found that the healthy human plasma B subgroup sEV has a remarkable cardiac function protection effect in an LPS-induced sepsis cardiomyopathy model, inflammatory factor expression can be reduced, cardiac function indexes can be improved, and a new treatment means is provided for treatment of sepsis cardiomyopathy.

The application belongs to the technical field of food deep processing, and particularly relates to a red date degraded polysaccharide, a degraded polysaccharide component, and a preparation method and application thereof. The application provides a preparation method of the red date degraded polysaccharide, in which the red date polysaccharide is degraded by using an ultrasonic wave assisted method combined with H2O2-Vc, and the red date degraded polysaccharide is separated and purified by using a DEAE-Sepharose Fast Flow gel column and a Sephacryl S-100 chromatographic column. The red date degraded polysaccharide and the polysaccharide component have the following characteristics: after the degradation treatment, the fragmentation degree of the polysaccharide surface is deepened, the degradation does not change the basic structure of the polysaccharide, and the apparent morphological characteristics change to a certain extent; the degradation treatment can improve the antioxidant capacity of the polysaccharide and is inversely proportional to the polysaccharide molecular weight.

The application discloses a method for separating and purifying low-molecular-weight polysaccharides from Polygonatum sibiricum. The method comprises the following steps: mixing the polysaccharides with fillers and / or filter aids, and then separating and purifying the polysaccharides. The fillers comprise Sephadex G-15 (cross-linked glucose gel), DEAE-Sepharose CL-4B (agarose gel) and Bio-Gel P-2 (polyacrylamide gel), and the filter aids comprise diatomite, perlite, cellulose, activated carbon and red clay. The method can significantly improve the yield of the low-molecular-weight polysaccharides with a molecular weight of less than 2 KDa, and the effect of the combination of the fillers and the filter aids is the best. The addition of the filter aids can avoid the blockage of the pores of the fillers in the middle and late stages of elution, improve the elution performance of the fillers on the polysaccharides and accelerate the elution and purification speed of the low-molecular-weight polysaccharides. The experiments show that the low-molecular-weight polysaccharides can significantly enhance the proliferation activity and phagocytosis of macrophages, promote the secretion of cytokines such as TNF-alpha, IL-6 and NO by RAW264.7 cells and improve the immune activity of the cells. The method has the characteristics of convenience, high efficiency and low cost and has high application value.