| 摘要 |
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. |
