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Thioredoxin 1 is responsible for antibody disulfide reduction in CHO cell culture |
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| Authors: | Koterba Kristen L Borgschulte Trissa Laird Michael W |
| Affiliation: | a Late Stage Cell Culture, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA b SAFC Biosciences, Sigma-Aldrich, 2909 Laclede Ave., St. Louis, MO 63103, USA |
| Abstract: | During large-scale manufacturing of an IgG1 monoclonal antibody in Chinese hamster ovary (CHO) cells, reduction of the antibody's disulfide bonds was observed. We present evidence that mammalian thioredoxin 1 (TXN1) is the terminal enzyme responsible for this reduction event. We demonstrate a marked prevention of IgG1 disulfide bond reduction in a cell-density dependent manner by knocking down expression of TXN1 via lentivirus transduction of short hairpin RNA. |
| Keywords: | mAb, monoclonal antibody IgG, immunoglobulin G CHO, Chinese hamster ovary TXN1, thioredoxin 1 TXN2, thioredoxin 2 GLRX, glutaredoxin NT, non-targeting gene NADPH, nicotinamide adenine dinucleotide phosphate shRNA, short-hairpin RNA |
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