Genetically modified cells and methods for expressing recombinant heparanase and methods of purifying same

Abstract

A method of purifying a recombinant heparanase from overexpressing cells or growth medium which comprises adsorbing said recombinant heparanase on an ion-exchange column under low salt conditions, washing said column with low salt solution thereby eluting other proteins, and eluting the recombinant heparanase from said column by a salt gradient or higher salt concentration.

What is claimed is:  
     
         1 . A method of purifying a recombinant heparanase from overexpressing cells or growth medium comprising adsorbing said recombinant heparanase on an ion-exchange column under low salt conditions, washing said column with low salt solution thereby eluting other proteins, and eluting the recombinant heparanase from said column by a salt gradient or higher salt concentration.  
     
     
         2 . The method of  claim 1 , wherein said ion-exchange column is a Source-S column.  
     
     
         3 . The method of  claim 1 , wherein said recombinant heparanase includes an amino acid sequence at least 70% homologous to SEQ ID NO:2.  
     
     
         4 . The method of  claim 1 , wherein said recombinant heparanase includes an amino acid sequence as set forth in SEQ ID NO:2.  
     
     
         5 . A method of affinity purifying heparanase comprising: 
 (a) contacting a heparanase preparation with an affinity substrate including a solid matrix and an anti-heparanase antibody being immobilized thereto, said anti-heparanase antibody being capable of specifically binding at least one epitope of said heparanase;    (b) washing said affinity substrate; and    (c) eluting heparanase molecules being adsorbed on said affinity substrate via said anti-heparanase antibody.    
     
     
         6 . The method of  claim 5 , wherein said heparanase includes an amino acid sequence at least 70% homologous to SEQ ID NO:2.  
     
     
         7 . The method of  claim 5 , wherein said heparanase includes an amino acid sequence as set forth in SEQ ID NO:2.