Patent ID: 6352852
Filing Date: 2002-03-05
Classification: C12N,C12Q,C12Y,Y10S

Abstract:
A method for the purification of human platelet heparanase, which method comprises the steps of:i. homogenizing human platelets in a CHAPS-containing solution to obtain heparanase-containing material; ii. contacting the heparanase material with a Concanavalin A-Sepharose chromatography matrix in the presence of Triton X-100 to bind heparanase activity to said matrix; iii. eluting a first purified heparanase-containing fraction from said matrix; iv. contacting said first purified heparanase-containing fraction with a Zn2+-chelating Sepharose chromatography matrix; v. contacting effluent from said Zn2+-chelating Sepharose chromatography matrix with a dye-resin matrix to bind heparanase activity to said dye-resin matrix, wherein said dye-resin matrix comprises a Blue A-resin or Reactive Red-resin matrix; vi. eluting a second purified heparanase-containing fraction from said dye-resin matrix; vii. contacting the second purified heparanase-containing fraction with an octyl-agarose matrix to remove contaminating proteins; and viii. purifying heparanase from effluent from said octyl-agarose matrix by gel filtration chromatography.