Process for manufacture of a modified collagen-induced platelet aggregation inhibitor pallidipin

US5882887A · kind A · utility

Assignee

• SCHERING CORPORATION · US

Inventors

• Christiane Noeske-Jungblut · Berlin, DE
• Andreas Becker · Celle, DE
• Bernard Haendler · Berlin, DE

Key dates

• Filing date: Mar 11, 1997
• Grant date: Mar 16, 1999
• Expiry date: Mar 11, 2017

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K2319/02
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

The invention provides a process of manufacture of a recombinant protein called Asp-Pallidipin. Asp-Pallidipin inhibits the collagen-induced platelet aggregation of mammalian platelets. The Asp-Pallidipin comprises PA1 (i) a protein (Pallidipin) selected from the group of Pallidipin proteins, and PA1 (ii) the amino acid aspartic acid, PA1 wherein the aspartic acid is connected by a peptide bond with the N-terminal end of Pallidipin. The process comprises the steps PA2 aa) transfecting at least one bacterium with an appropriate vector, wherein the vector comprises: PA3 (i) a DNA or cDNA coding for the recombinant Asp-Pallidipin PA3 (ii) a suitable signal peptide sequence which signal sequence is cleaved so that the amino acid aspartic acid is in the position +1 of the amino acid sequence seen from the position of the Pallidipin and PA3 (iii) a suitable promoter; PA2 bb) expressing the preprotein comprising Asp-Pallidipin and the signal sequence; PA2 cc) transporting the Asp-Pallidipin from the cytoplasm of the bacterium to the periplasm, PA2 cleavage of the preprotein by at least one protease during the transport, producing the Asp-Pallidipin, PA2 dd) isolating the Asp-Pallidipin by…