Preparation method of low molecular weight heparin

Abstract

A preparation method of low molecular weight heparin comprises the following steps: a) cross-linking Phe-Leu-Asn-Gln-Asp and sepharose; b) adding the cross-linked gel to a 46 mM sodium phosphate buffer solution with the volume 8 times that of the gel, performing swelling, and sucking the obtained supernatant; c) dissolving heparin in the buffer solution with the volume 1 time that of the heparin,performing sample loading according to a ratio of the gel to the heparin of 0.5 mg: 3 mg, stirring the gel and heparin by using a stirring rod for 3 min, standing the obtained solution for 15 min, andsucking the obtained supernatant; d) adsorbing the active central site of the heparin to the pentapeptide of the gel, adding heparinase, and carrying out stirring hydrolysis at 35 DEG C in a water bath; e) randomly partially hydrolyzing the free part of the heparin, centrifuging the obtained solution, and taking the obtained precipitate; f) adding a 1M NaCl solution with the volume 1 time that ofthe precipitate to the precipitate, centrifuging the obtained solution, and taking the obtained supernatant; and g) concentrating the supernatant, allowing the concentrated supernatant to go througha purified water bio gel P2 column, performing automatic partial collection, performing 220 nm ultraviolet detection, removing salts, collecting obtained ultraviolet absorption parts, mixing the collected parts, and freeze-drying the obtained mixture.

Description

technical field [0001] The invention relates to a preparation method of low molecular weight heparin, which belongs to the technical field of biochemistry. Background technique [0002] The anticoagulant activity of heparin depends on its interaction with the serine protease inhibitor, antithrombin III (AT). During binding to heparin, AT undergoes a conformational change that enhances its irreversible ability to inhibit thrombin, including thrombin and factor Xa. Heparin-AT mutual recognition is an example of a well-studied polysaccharide-protein interaction. The recognition of heparin and AT is dependent on specific heparin pentasaccharide domains in heparin. The main structural feature of this specific heparin pentasaccharide is that there is a 3-O sulfate group modification at the glucosamine residue in the middle of the pentasaccharide. The three-position sulfated modified fragments of low molecular weight heparin are reduced, and the molecular weight is reduced, so t...

AI Technical Summary

Problems solved by technology

Low-molecular-weight heparin has reduced three-position sulfated fragments and reduced molecular weight, so the activity is reduced, but the safety is increased