title: A rapid method for simultaneous evaluation of free light chain content and aggregate content in culture media of Chinese hamster ovary cells expressing monoclonal antibodies for cell line screening.
abstract: The goal of developing a monoclonal antibody (mAb) production process is high productivity and high quality. Because the productivity and quality of mAbs depend on cell line properties, the selection of cell lines suitable for large-scale production is an important stage in process development for mAb production. The light chain (LC) is important for antibody folding and assembly in the endoplasmic reticulum; cell lines that secrete a large amount of LCs in the medium secrete high-quality antibodies with high productivity. LC contents in culture media have been estimated by western blotting, reverse-phase high-performance liquid chromatography, and enzyme-linked immunosorbent assay. However, these analyses require fine tuning of experimental conditions for each antibody analyzed. Here we report a rapid and simple high-sensitivity size-exclusion chromatography (HS-SEC) method to evaluate the contents of low-molecular weight species (LMWS, mainly consisting of LC monomers and dimers) and high-molecular weight species (HMWS, aggregates) in the media for cell line screening. Because LMWS and HMWS are important indicators of productivity and quality, respectively, for cell line screening, HS-SEC will be useful in the first step of cell line selection needed for large-scale production.
authors.affiliation: Bio-process Research and Development Laboratories, Kyowa Hakko Kirin Co., Ltd., 100-1 Hagiwara-machi, Takasaki-shi, Gunma 370-0013, Japan; Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu-shi, Gunma 376-8515, Japan. Electronic address: yoichi.ishii@kyowa-kirin.co.jp.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Ishii
authors.affiliation: Kyowa Hakko Kirin Co., Ltd., 1-6-1 Ohte-machi, Chiyoda-ku, Tokyo 100-8185, Japan.
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Tsukahara
authors.affiliation: Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu-shi, Gunma 376-8515, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Wakamatsu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D016466:CHO Cells; D002850:Chromatography, Gel / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D007147:Immunoglobulin Light Chains / Q000032:analysis* / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D008970:Molecular Weight; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D003470:Culture Media; D007147:Immunoglobulin Light Chains
keywords: High-molecular weight species; Low-molecular weight species; Size-exclusion chromatography; Therapeutic antibody