UNASSIGNED: Antibody @entity1520 bond reduction during monoclonal antibody (mAb) production is a phenomenon that has been attributed to the reducing enzymes from CHO cells acting on the mAb during the harvest process. However, the impact of antibody reduction on the downstream purification process has not been studied. During the production of an IgG2 mAb, antibody reduction was observed in the harvested cell culture fluid (HCCF), resulting in high fragment levels. In addition, aggregate levels increased during the low pH treatment step in the purification process. A correlation between the level of free @entity1432 in the HCCF (as a result of antibody reduction) and aggregation during the low pH step was established, wherein higher levels of free @entity1432 in the starting sample resulted in increased levels of aggregates during low pH treatment. The elevated levels of free @entity1432 were not reduced over the course of purification, resulting in carry-over of high free @entity1432 content into the formulated drug substance. When the drug substance with high free @entity1433 was monitored for product degradation at room temperature and 2   -   8  C, faster rates of aggregation were observed compared to the drug substance generated from HCCF that was purified immediately after harvest. Further, when antibody reduction mitigations (e.g. chilling, aeration and addition of @entity1434 ) were applied, HCCF could be held for an extended period of time while providing the same product quality/stability as material that had been purified immediately after harvest. This article is protected by copyright. All rights reserved.

Effects of antibody XXXX bond reduction on purification process performance and final drug substance stability.

@entity1432 :: ('CHEBI:29256', 'Chemical') :: ['thiol'] @entity1433 :: ('MESH:D013438', 'Chemical') :: ['thiols'] @entity1434 :: ('MESH:D003553', 'Chemical') :: ['cystine'] @entity1520 :: ('CHEBI:48343', 'Chemical') :: ['disulfide']
@entity1520 :: (CHEBI:48343,Chemical) :: ['disulfide']
