Identification and prevention of antibody disulfide bond reduction during cell culture manufacturing
Article first published online: 22 FEB 2010
Copyright © 2010 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 106, Issue 3, pages 452–461, 15 June 2010
How to Cite
Trexler-Schmidt, M., Sargis, S., Chiu, J., Sze-Khoo, S., Mun, M., Kao, Y.-H. and Laird, M. W. (2010), Identification and prevention of antibody disulfide bond reduction during cell culture manufacturing. Biotechnol. Bioeng., 106: 452–461. doi: 10.1002/bit.22699
- Issue published online: 23 APR 2010
- Article first published online: 22 FEB 2010
- Accepted manuscript online: 22 FEB 2010 12:00AM EST
- Manuscript Accepted: 8 FEB 2010
- Manuscript Revised: 23 DEC 2009
- Manuscript Received: 31 JUL 2009
In the biopharmaceutical industry, therapeutic monoclonal antibodies are primarily produced in mammalian cell culture systems. During the scale-up of a monoclonal antibody production process, we observed excessive mechanical cell shear as well as significant reduction of the antibody's interchain disulfide bonds during harvest operations. This antibody reduction event was catastrophic as the product failed to meet the drug substance specifications and the bulk product was lost. Subsequent laboratory studies have demonstrated that cells subjected to mechanical shear release cellular enzymes that contribute to this antibody reduction phenomenon (manuscript submitted; Kao et al., 2009). Several methods to prevent this antibody reduction event were developed using a lab-scale model to reproduce the lysis and reduction events. These methods included modifications to the cell culture media with chemicals (e.g., cupric sulfate (CuSO4)), pre- and post-harvest chemical additions to the cell culture fluid (CCF) (e.g., CuSO4, EDTA, L-cystine), as well as lowering the pH and air sparging of the harvested CCF (HCCF). These methods were evaluated for their effectiveness in preventing disulfide bond reduction and their impact to product quality. Effective prevention methods, which yielded acceptable product quality were evaluated for their potential to be implemented at manufacturing-scale. The work described here identifies numerous effective reduction prevention measures from lab-scale studies; several of these methods were then successfully translated into manufacturing processes. Biotechnol. Bioeng. 2010; 106: 452–461. © 2010 Wiley Periodicals, Inc.