PURIFYING OF TRYPSIN AND TRYPSIN LIKE ENZYME AND STORING OF PURIFIED ENZYME

摘要

1388725 Enzyme attachable complexes XEROX CORP 11 May 1972 [12 May 1971] 22057/72 Headings C3U C3A C3P and C3H An affinity matrix material, used to adsorb and purify trypsin or trypsin-like enzymes comprises an insoluble matrix and a ligand, said ligand having the formula wherein the insoluble matrix may be cellulose, aminoethyl cellulose, phospho cellulose, DEAE cellulose, CMC, ECTEOLA cellulose, p-aminobenzyl cellulose, polyethyleneiminocellulose, triethylamino cellulose, sulphoethyl cellulose, guanidoethyl cellulose, agaroses, crosslinked dextrans, crosslinked polyacrylamides, agaropectin, or crosslinked or non crosslinked collagen. A is a divalent aliphatic, cycloaliphatic, aryl or arylaliphatic group, preferably a pentylene, hexylene, norvalylene, cyclohexylene, phenylene or carboxyphenylene radical and the preferred ligands are m-aminobenzamidine, paminobenzamidine or p-aminophenylguanidine. The ligand is preferably attached to the matrix by an intermediate leash molecule, e.g. epsilon aminocaproic acid; betaalanine; 1,6-diaminohexane; bis-(3-aminopropyl) amine; glycine; glycylglycine, glycylglycylglycine; succinamidoethylamine; succinamidobutylamine; succinamidohexamethylamine; aminoethanol; or 2,2SP1/SP- diaminodiethyl ether. Trypsin or trypsin-like enzymes may be purified by contacting a mixture of the enzyme with the affinity matrix material to adsorb the enzyme, washing the matrix material after the adsorption and then eluting the enzyme with a liquid comprising a competitive inhibitor material, e.g. benzamidine hydrochloride. Enzymes may be trypsin, carboxypeptidase B, papain, ficin, thrombin, plasmin, subtilopeptidase, aspergillopeptidase, streptococcuspeptidase, clostridiopeptidase, bromelain or urokinase.