Untargeted proteomics reveals upregulation of stress response pathways during CHO-based monoclonal antibody manufacturing process leading to disulfide bond reduction

mAbs

卷:13

Zhengjian Li¹  Susan Egan¹  Sanchayita Ghose¹  Mengyuan Zheng¹  Michael Borys¹  Anthony J. Cura¹  Kathryn L. Aron¹  Xuankuo Xu¹  Seo-Young Park²  Kyongbum Lee² 

[1] Biologics Development, Global Product Development and Supply, Bristol-Myers Squibb, Devens, USA;

[2] Department of Chemical and Biological Engineering, Tufts University, Medford, MA, USA;

关键词: Monoclonal antibody;    disulfide bond reduction;    CHO cell culture;    proteomics;    heat shock proteins;    cellular stress response;   

DOI  :  10.1080/19420862.2021.1963094

来源: DOAJ

【 摘 要 】

Monoclonal antibody (mAb) interchain disulfide bond reduction can cause a loss of function and negatively impact the therapeutic’s efficacy and safety. Disulfide bond reduction has been observed at various stages during the manufacturing process, including processing of the harvested material. The factors and mechanisms driving this phenomenon are not fully understood. In this study, we examined the host cell proteome as a potential factor affecting the susceptibility of a mAb to disulfide bond reduction in the harvested cell culture fluid (HCCF). We used untargeted liquid-chromatography-mass spectrometry-based proteomics experiments in conjunction with a semi-automated protein identification workflow to systematically compare Chinese hamster ovary (CHO) cell protein abundances between bioreactor conditions that result in reduction-susceptible and reduction-free HCCF. Although the growth profiles and antibody titers of these two bioreactor conditions were indistinguishable, we observed broad differences in host cell protein (HCP) expression. We found significant differences in the abundance of glycolytic enzymes, key protein reductases, and antioxidant defense enzymes. Multivariate analysis of the proteomics data determined that upregulation of stress-inducible endoplasmic reticulum (ER) and other chaperone proteins is a discriminatory characteristic of reduction-susceptible HCP profiles. Overall, these results suggest that stress response pathways activated during bioreactor culture increase the reduction-susceptibility of HCCF. Consequently, these pathways could be valuable targets for optimizing culture conditions to improve protein quality.

【 授权许可】

Unknown