Effect of the disulfide isomerase PDIa4 on the antibody production of Chinese hamster ovary cells

• Published in: Journal of bioscience and bioengineering 2020-12, Vol.130 (6), p.637-643
• Main Authors: Komatsu, Kei, Kumon, Kento, Arita, Mayuno, Onitsuka, Masayoshi, Omasa, Takeshi, Yohda, Masafumi
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - biosynthesis; Antibody; Chaperone; Chinese hamster ovary; CHO Cells; Cricetinae; Cricetulus; Folding; Gene Expression Regulation, Enzymologic; PDIa4; Protein disulfide-isomerase; Protein Disulfide-Isomerases - genetics; Protein Disulfide-Isomerases - metabolism; RNA, Messenger - genetics
• ISSN: 1389-1723; 1347-4421
• DOI: 10.1016/j.jbiosc.2020.08.001

Therapeutic monoclonal antibodies recognize and bind specific molecules on the surface of target cells, stimulating the immune system, which can attack these targeted cells. These antibodies are produced by mammalian cells, including Chinese hamster ovary (CHO) cells, because the formation of antibodies requires complicated posttranslational modifications, including peptidyl–prolyl cis/trans isomerization, disulfide bond formation, and glycosylation. Currently, it is thought that the efficient production of secretory proteins is limited by posttranslational processes. The ER is the biosynthesis site of all secreted and membrane proteins. The accumulation of unfolded proteins in the ER causes the ER stress response. During the ER stress state, various molecular chaperones are expressed to prevent proteins from the aggregate formation. The molecular chaperone involved in ER stress likely plays an essential role in the production of secretory proteins. The purpose of this study was to improve the production of monoclonal antibodies by cells. We elucidated the function of ER chaperones in the production of a monoclonal antibody. First, we quantitatively measured the mRNA expression levels of protein disulfide-isomerase family members. In CHO HcD6 cells treated with tunicamycin, the expression level of pdia4 was significantly increased. Second, we investigated the relationship between PDIa4 and antibody productivity in pdia4-knockdown cells. Both a decrease in the amount of secreted antibody and the accumulation of immature antibodies inside the cells were observed. Recombinant PDIa4 was able to refold the antibodies and Fabs. These results indicate that PDIa4 affects the production of monoclonal antibodies by catalyzing disulfide bond formation in these antibodies in CHO cells.