title: Effect of temperature shift on levels of acidic charge variants in IgG monoclonal antibodies in Chinese hamster ovary cell culture.
abstract: During the production of therapeutic monoclonal antibodies (mAbs), not only enhancement of mAb productivity but also control of quality attributes is critical. Charge variants, which are among the most important quality attributes, can substantially affect the in vitro and in vivo properties of mAbs. During process development for the production of mAbs in a Chinese hamster ovary cell line, we have observed that an improvement in mAb titer is accompanied by an increase in the content of acidic charge variants. Here, to help maintain comparability among mAbs, we aimed to identify the process parameters that controlled the content of acidic charge variants. First, we used a Plackett-Burman design to identify the effect of selected process parameters on the acidic charge variant content. Eight process parameters were selected by using a failure modes and effects analysis. Among these, temperature shift was identified from the Plackett-Burman design as the factor most influencing the acidic charge variant content. We then investigated in more detail the effects of shift temperature and temperature shift timing on this content. The content decreased with a shift to a lower temperature and with earlier timing of this temperature shift. Our observations suggest that Plackett-Burman designs are advantageous for preliminary screening of bioprocess parameters. We report here for the first time that temperature downshift is beneficial for effective control of the acidic peak variant content.
authors.affiliation: Project Planning and Coordination Department, Project and Lifecycle Management Unit, Chugai Pharmaceutical Co., Ltd., 1-1 Nihonbashi-Muromachi 2-Chome, Chuo-ku, Tokyo 103-8324, Japan; Life Science and Bioengineering, Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1 Tennodai 1-Chome, Tsukuba, Ibaraki 305-8572, Japan. Electronic address: kishishitasuh@chugai-pharm.co.jp.
authors.forename: Shohei
authors.initials: S
authors.lastname: Kishishita
authors.affiliation: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Nishikawa
authors.affiliation: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Shinoda
authors.affiliation: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nagashima
authors.affiliation: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Okamoto
authors.affiliation: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Takuma
authors.affiliation: Life Science and Bioengineering, Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1 Tennodai 1-Chome, Tsukuba, Ibaraki 305-8572, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Aoyagi
mesh_terms: D000143:Acids / Q000737:chemistry*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000737:chemistry*; D013696:Temperature
chemical_list: D000143:Acids; D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G
keywords: Acidic charge variants; Chinese hamster ovary cells; IgG monoclonal antibody; Plackett–Burman design; Temperature shift