Culture temperature modulates aggregation of recombinant antibody in cho cells
Article first published online: 3 OCT 2011
Copyright © 2011 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 109, Issue 1, pages 125–136, January 2012
How to Cite
Gomez, N., Subramanian, J., Ouyang, J., Nguyen, M. D.H., Hutchinson, M., Sharma, V. K., Lin, A. A. and Yuk, I. H. (2012), Culture temperature modulates aggregation of recombinant antibody in cho cells. Biotechnol. Bioeng., 109: 125–136. doi: 10.1002/bit.23288
- Issue published online: 17 NOV 2011
- Article first published online: 3 OCT 2011
- Accepted manuscript online: 30 SEP 2011 06:49AM EST
- Manuscript Accepted: 26 JUL 2011
- Manuscript Revised: 16 JUL 2011
- Manuscript Received: 11 APR 2011
- light chain;
- heavy chain
During production of therapeutic monoclonal antibodies (mAb), it is highly desirable to remove and control antibody aggregates in the manufacturing process to minimize the potential risk of immunogenicity to patients. During process development for the production of a recombinant IgG in a CHO cell line, we observed atypical high variability from 1 to 20% mAb aggregates formed during cell culture that negatively impacted antibody purification. Analytical characterization revealed the IgG aggregates were mediated by hydrophobic interactions likely caused by misfolded antibody during intracellular processing. Strikingly, data analysis showed an inverse correlation of lower cell culture temperature producing higher aggregate levels. All cultures at 37°C exhibited ≤5% aggregates at harvest. Aggregate levels increased 4–12-fold in 33°C cultures when compared to 37°C, with a corresponding 2–4-fold increase in heavy chain (HC) and light chain (LC) mRNA. Additionally, 37°C cases showed a greater excess of LC to HC mRNA levels. Endoplasmic reticulum (ER) chaperone expression and ER size also increased 25–75% at 33°C versus 37°C but to a lesser extent than LC and HC mRNA, consistent with a potential limiting ER folding capacity at 33°C for this cell line. Finally, we identified a 2–5-fold increase in mAb aggregate formation at 33°C compared to 37°C cultures for three additional CHO cell lines. Taken together, our observations indicate that low culture temperature can increase antibody aggregate formation in CHO cells by increasing LC and HC transcripts coupled with limited ER machinery. Biotechnol. Bioeng. 2012;109: 125–136. © 2011 Wiley Periodicals, Inc.