Title:
【発明の名称】精製方法
Document Type and Number:
Japanese Patent JPH01500563
Kind Code:
A
Abstract:
Process for the purification of tPA comprising: applying a mother solution containing tPA to a highly anionic chromatography medium (HACM) equilibrated with a buffer at 25mS ionic strength or lower and at a pH between 4 and 7; eluting the tPA from the highly anionic chromatography medium with a high ionic strength buffer; applying the eluate from the highly anionic chromatography medium to an affinity chromatography medium (ACM), comprising an anti-tPA antibody covalently bound to a chromatography material, equilibrated with a buffer at pH 5 to 10; and eluting the tPA from the affinity chromatography medium with a buffer which disrupts the binding of the antibody to the tPA. Preferably, the mother solution is the supernatant from a culture of a eukaryotic cell line which has been transformed by recombinant DNA technology so that it is able to express and secrete tPA. Advantageously the mother solution is processed as soon as it is harvested and all the process steps are carried out as quickly as possible, the mother solution contains a serine protease inhibitor, and all the solutions contain a detergent and are kept at as low a pH as possible.
Inventors:
Kennie, Andrew Charles
Angela Joyce, Pery
White, susan
Marston, Fiona Angela Olinda
Angela Joyce, Pery
White, susan
Marston, Fiona Angela Olinda
Application Number:
JP50435687A
Publication Date:
March 01, 1989
Filing Date:
July 15, 1987
Export Citation:
Assignee:
Celtec Limited
International Classes:
C12N9/64; A61K38/00; A61K38/46; A61P7/02; C07K1/22; C07K14/00; C07K14/81; C07K16/00; C12N9/72; C12P21/00; C12R1/91; (IPC1-7): A61K37/54; C07K3/18; C07K13/00; C12N9/64; C12P21/00
Attorney, Agent or Firm:
Akira Asamura (2 outside)