BIOT 440 |
| The purification of monoclonal antibodies typically involves multiple chromatography steps exploiting orthogonal modes of separation. These chromatography steps use resins consisting of a support matrix onto which particular functional chemistries are immobilized. Using polyelectrolytes, it may be possible to exploit these same functional chemistries to achieve protein purification in solution. Through the manipulation of solution pH and ionic strength, an antibody-polyelectrolyte complex can form leading to antibody precipitation. Purification occurs through selective partitioning of the antibody and impurities into the solid or liquid phases. The potential for polyelectrolyte induced precipitation of antibodies to replace traditional chromatography steps was evaluated using polyvinylsulfonic acid (PVS). PVS precipitation was evaluated as a replacement for the initial capture step as well as an intermediate polishing step in the purification of a humanized IgG1 monoclonal antibody. PVS precipitation separated the antibody from host cell proteins, leached protein A, small molecules such as insulin and gentamicin as well as antibody fragments and aggregates. PVS was subsequently removed from antibody pools to < 1ug/mL using anion exchange chromatography. PVS precipitation did not impact the biological activity of the re-suspended antibody. |
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Downstream Processing: Non-Chromatographic Separation Techniques: Improving Process Throughput
8:00 AM-11:00 AM, Thursday, August 23, 2007 BCEC -- 107B, Oral
Division of Biochemical Technology |