title: Thioredoxin 1 is responsible for antibody disulfide reduction in CHO cell culture.
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.
authors.affiliation: Late Stage Cell Culture, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA. koterba.kristen@gene.com
authors.forename: Kristen L
authors.initials: KL
authors.lastname: Koterba
authors.forename: Trissa
authors.initials: T
authors.lastname: Borgschulte
authors.forename: Michael W
authors.initials: MW
authors.lastname: Laird
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D016466:CHO Cells / Q000378:metabolism; D049109:Cell Proliferation; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D004220:Disulfides / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D015202:Protein Engineering / Q000379:methods; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D013879:Thioredoxins / Q000235:genetics*; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D004220:Disulfides; D007074:Immunoglobulin G; D034741:RNA, Small Interfering; D013879:Thioredoxins