title: Optimization of chemically defined feed media for monoclonal antibody production in Chinese hamster ovary cells.
abstract: Chinese hamster ovary (CHO) cells are the most commonly used mammalian host for large-scale commercial production of therapeutic monoclonal antibodies (mAbs). Chemically defined media are currently used for CHO cell-based mAb production. An adequate supply of nutrients, especially specific amino acids, is required for cell growth and mAb production, and chemically defined fed-batch processes that support rapid cell growth, high cell density, and high levels of mAb production is still challenging. Many studies have highlighted the benefits of various media designs, supplements, and feed addition strategies in cell cultures. In the present study, we used a strategy involving optimization of a chemically defined feed medium to improve mAb production. Amino acids that were consumed in substantial amounts during a control culture were added to the feed medium as supplements. Supplementation was controlled to minimize accumulation of waste products such as lactate and ammonia. In addition, we evaluated supplementation with tyrosine, which has poor solubility, in the form of a dipeptide or tripeptide to improve its solubility. Supplementation with serine, cysteine, and tyrosine enhanced mAb production, cell viability, and metabolic profiles. A cysteine-tyrosine-serine tripeptide showed high solubility and produced beneficial effects similar to those observed with the free amino acids and with a dipeptide in improving mAb titers and metabolic profiles.
authors: Project Planning & Coordination Department, Project & 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: Shohei
authors: S
authors: Kishishita
authors: Human Resource Management Department, Chugai Pharmaceutical Co., Ltd., 1-1 Nihonbashi-Muromachi 2-Chome, Chuo-ku, Tokyo 103-8324, Japan.
authors: Satoshi
authors: S
authors: Katayama
authors: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors: Kunihiko
authors: K
authors: Kodaira
authors: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors: Yoshinori
authors: Y
authors: Takagi
authors: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors: Hiroki
authors: H
authors: Matsuda
authors: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors: Hiroshi
authors: H
authors: Okamoto
authors: API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
authors: Shinya
authors: S
authors: Takuma
authors: Utsunomiya Plant Manufacturing, Chugai Pharmaceutical Manufacturing Co., Ltd., 16-3 Kiyohara Kogyo Danchi, Utsunomiya, Tochigi 321-3231, Japan.
authors: Chikashi
authors: C
authors: Hirashima
authors: 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: Hideki
authors: H
authors: Aoyagi
mesh_terms: D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology; D000641:Ammonia / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation / Q000187:drug effects*; D016466:CHO Cells; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D019344:Lactic Acid / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism / Q000494:pharmacology; D012995:Solubility
chemical_list: D000596:Amino Acids; D000911:Antibodies, Monoclonal; D003470:Culture Media; D009842:Oligopeptides; D019344:Lactic Acid; D000641:Ammonia
keywords: Amino acid; Chemically defined medium; Chinese hamster ovary cells; Monoclonal antibody; Tripeptide