title: Overexpression of CHOP alone and in combination with chaperones is effective in improving antibody production in mammalian cells.
abstract: Secretory capacities including folding and assembly are believed to be limiting factors in the establishment of mammalian cell lines producing high levels of recombinant therapeutic proteins. To achieve industrial success, it is also important to improve protein folding, assembly, and secretory processes in combination with increasing transcription and translation. Here, we identified the expression of CHOP/Gadd153 and GRP78, which are unfolded protein response (UPR)-related genes, correlated with recombinant antibody production in stable CHO cells. Subsequently, CHOP overexpression resulted in increasing recombinant antibody production in some mammalian cell lines, and in addition a threefold further enhancement was obtained by combining expression with UPR-related genes or ER chaperones in transient assays. Overexpression of CHOP had no effect on the biochemical characteristics of the product. These results suggest overexpression of CHOP and its combinations may be an effective method to efficiently select a single cell line with a high level of antibody production in the development of cell lines for manufacturing.
authors.affiliation: R&D Division, Biologics Research Laboratories, Daiichisankyo Co., Ltd., 1-2-58 Hiromachi, Shinagawa-ku, Tokyo 140-8710, Japan. nishimiya.daisuke.tc@daiichisankyo.co.jp
authors.forename: Daisuke
authors.initials: D
authors.lastname: Nishimiya
authors.forename: Takashi
authors.initials: T
authors.lastname: Mano
authors.forename: Kenji
authors.initials: K
authors.lastname: Miyadai
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yoshida
authors.forename: Tohru
authors.initials: T
authors.lastname: Takahashi
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D051742:Transcription Factor CHOP / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000906:Antibodies; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D051742:Transcription Factor CHOP