Method for the production of FVII

Abstract

Disclosed is a method for high efficiency release of recombinant proteins in eukaryotic cells and more specifically, for enhancing the secretion of Factor VII by co-expression of kex2 endoprotease with FVII in cells of mammalian origin.

What is claimed is:  
     
         1 . A method for producing Factor VII comprising (a) cultivation of a mammalian cell line comprising a DNA sequence encoding a yeast endoprotease or a derivative thereof and a DNA sequence encoding FVII in a suitable culture medium; and (b) isolation of Factor VII from the medium.  
     
     
         2 . The method of  claim 1 , wherein the yeast endoprotease is derived from a Saccharomyces strain.  
     
     
         3 . The method if  claim 2 , wherein the yeast endoprotease is a Kex2 like endoprotease.  
     
     
         4 . The method of  claim 3  wherein the yeast Kex2 like endoprotease is C-terminally truncated and has no transmembrane region.  
     
     
         5 . The method of  claim 4 , wherein the yeast Kex2 like endoprotease has ER retention signal added to the C-terminal end.  
     
     
         6 . The method of  claim 2  wherein the yeast endoprotease is the Kex2 endoprotease from  Saccharomyces cerevisiae.    
     
     
         7 . The method of  claim 1 , wherein the culture medium is a serum free medium.  
     
     
         8 . The method of  claim 1  wherein the mammalian cell line is a CHO cell line.  
     
     
         9 . The method of  claim 1  wherein the mammalian cell line is a BHK cell line.  
     
     
         10 . The method of  claim 1  wherein the mammalian cell line is a HEK 293 cell line.