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Culture temperature modulates aggregation of recombinant antibody in cho cells
Author(s) -
Gomez Natalia,
Subramanian Jayashree,
Ouyang Jun,
Nguyen Mary D.H.,
Hutchinson Matthew,
Sharma Vikas K.,
Lin Andy A.,
Yuk Inn H.
Publication year - 2012
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.23288
Subject(s) - chinese hamster ovary cell , endoplasmic reticulum , intracellular , antibody , cell culture , monoclonal antibody , recombinant dna , immunogenicity , chemistry , messenger rna , microbiology and biotechnology , biochemistry , biology , immunology , gene , receptor , genetics
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.