Heparanase derived from human Sk-Hep-1 cell line
AssigneeHadassah Medical Organization Kiryat Hadassah
InventorsZvi Fuks, Israel Vlodavsky
Life of this patent
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Abstract
The present invention relates to a substantially purified heparanase obtained from the human SK-HEP-1 cell line and a method to purify the heparanase from a centrifuged dialyzed homogenate of the SK-HEP-1 cells or a cell culture medium containing the heparanase from the SK-HEP-1 cell line.
Claims39
A heparanase formulation, obtained from the Sk-Hep-1 cell line wherein the heparanase has hydrolytic activity for heparan sulfate but not heparin and the specific heparanase activity of the formulation is at least 1000-fold greater than the specific heparanase activity of a centrifuged dialyzed cell homogenate prepared from the Sk-Hep-1 cell line.
A heparanase formulation obtained from the human SK-HEP-1 cell line wherein the heparanase has hydrolytic activity for heparan sulfate but not heparin, the specific heparanase activity of the formulation is at least 1000-fold greater than the specific heparanase activity of a centrifuged dialyzed cell homogenate prepared from the SK-HEP-1 cell line, and the heparanase is obtained by the method comprising:
- (a) contacting a heparanase-containing centrifuged dialyzed homogenate Of cell line Sk-Hep-1 with a cation exchange resin, and obtaining a first active eluate;
- (b) contacting the first eluate with an affinity purification absorbent;
- and
- (c) recovering a second active eluate which contains heparanase to provide for said heparanase formulation.
A method for preparing a heparanase formulation, which comprises:
- (a) obtaining a heparanase containing cell supernatant from the human Sk-HEP-1 line;
- (b) contacting the heparanase containing supernatant with a cation exchange resin, and obtaining a first active eluate;
- (c) contacting the first eluate with an affinity purification absorbent;
- and
- (d) recovering a second active eluate which contains heparanase to provide for said heparanase formulation.
A heparanase formulation obtained from the human SK-HEP-1 cell line wherein the heparanase has hydrolytic activity for heparan sulfate but not heparin, the specific heparanase activity of the formulation is at least 1000-fold greater than the specific heparanase activity of a centrifuged dialyzed cell homogenate prepared from the SK-HEP-1 cell line, and the heparanase is obtained by the method comprising:
- (a) obtaining a heparanase-containing human SK-HEP-1 cell line culture medium;
- (b) contacting the heparanase containing medium with a cation exchange resin, and obtaining a first active eluate;
- (c) contacting the first eluate with an affinity purification absorbent;
- and
- (d) recovering a second active eluate which contains heparanase to provide for said heparanase formulation, wherein the heparanase is present in Sk-Hep-1.
Dependencies are inferred from the claim text.