Journal article
BiP Inducer X: An ER Stress Inhibitor for Enhancing Recombinant Antibody Production in CHO Cell Culture
CHO Cell Line Engineering and Design, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
CHO in Silico Engineering of Glycosylation and Protein Quality (CiSe), Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
CHO Core, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark4
Prolonged endoplasmic reticulum (ER) stress reduces protein synthesis and induces apoptosis in mammalian cells. When dimethyl sulfoxide (DMSO), a specific monoclonal antibody productivity (qmAb )-enhancing reagent, is added to recombinant Chinese hamster ovary (rCHO) cell cultures (GSR cell line), it induces ER stress and apoptosis in a dose-dependent manner.
To determine an effective ER stress inhibitor, three ER stress inhibitors (BiP inducer X [BIX], tauroursodeoxycholic acid, and carbazole) are examined and BIX shows the best production performance. Coaddition of BIX (50 μm) with DMSO extends the culture longevity and enhances qmAb . As a result, the maximum mAb concentration is significantly increased with improved galactosylation.
Coaddition of BIX significantly increases the expression level of binding immunoglobulin protein (BiP) followed by increased expression of chaperones (calnexin and GRP94) and galactosyltransferase. Furthermore, the expression levels of CHOP, a well-known ER stress marker, and cleaved caspase-3 are significantly reduced, suggesting that BIX addition reduces ER stress-induced cell death by relieving ER stress.
The beneficial effect of BIX on mAb production is also demonstrated with another qmAb -enhancing reagent (sodium butyrate) and a different rCHO cell line (CS13-1.00). Taken together, BIX is an effective ER stress inhibitor that can be used to increase mAb production in rCHO cells.
Language: | English |
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Year: | 2019 |
Pages: | e1900130 |
ISSN: | 18607314 and 18606768 |
Types: | Journal article |
DOI: | 10.1002/biot.201900130 |
ORCIDs: | 0000-0002-9876-7378 , Kwang Ha, Tae , Hansen, Anders H and Kildegaard, Helene Faustrup |
BiP inducer X Chinese hamster ovary (CHO) cells endoplasmic reticulum (ER) stress inhibitors galactosylation monoclonal antibodies
1-(3,4-dihydroxyphenyl)-2-thiocyanate-ethanone Animals Antibodies, Monoclonal Apoptosis Batch Cell Culture Techniques CHO Cells Cricetulus Dimethyl Sulfoxide Endoplasmic Reticulum Chaperone BiP Endoplasmic Reticulum Stress Galactose Heat-Shock Proteins Recombinant Proteins Rituximab Thiocyanates