Expression of heparin-binding protein in recombinant mammalian cells

Abstract

The present invention relates to a method for producing a mammalian heparin-binding protein in a mammalian cell that can be cultured under anaerobic conditions after introducing a nucleic acid encoding as heparin-binding protein into said cell with comprising (a) introducing into said mammalian cell a nucleic acid encoding said heparin-binding protein; (b) culturing the cell of step (a) under conditions conducive to expression of said HBP; and (c) recovering said HBP from the culture medium. Furthermore, the invention relates to the recombinant mammalian cell used in the method.

Description

field of invention

[0001] The present invention relates to a method for producing heparin-binding protein (HBP) in recombinant mammalian cells cultureable under anaerobic conditions after transfection with a nucleic acid encoding a heparin-binding protein. The invention also relates to said recombinant mammalian cells. Background of the invention

[0002] The covalent structures of two closely related proteins isolated from peripheral blood neutrophils of human and porcine origin have recently been determined (see H. Flodgaard et al., 1991, Eur. J. Biochem. 197: 535-547; J. Pohl et al., 1990, FEBS Lett. 272:200 et al.). These two proteins have a high degree of similarity to neutrophil elastase, but by selectively incorporating the active serine at position 195 and the active histidine at position 57 (chymotrypsin numbering (B.S.Hartley, 1970, Phil .Trans.Roy.Soc.Series 257:77 ff)) After the mutation, its protease activity was lost. These two protei...