title: Electrostatic engineering of the interface between heavy and light chains promotes antibody Fab fragment production.
abstract: Monoclonal antibodies and antibody fragments are used for diverse diagnostic and therapeutic applications. We have investigated the secretory production of Fab fragments from insect cells cotransfected with plasmid vectors carrying heavy- and light-chain genes. In the present study, to promote the formation of the disulfide bond between the heavy and light chains, some positively charged amino acid residues were introduced near the cysteine residue for the disulfide bond at the C-terminus of CL, while some negatively charged amino acid residues were added near the cysteine residue for the disulfide bond at the C-terminus of CH1. This electrostatic steering led to an increase in Fab secretions from insect cells.
authors: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.|Manufacturing Technology Association of Biologics (MAB), c/o Center for Business Incubation, Kobe University, 1-5-6 Minatojima-Minamimachi, Chuo-ku, Kobe, 650-0047, Japan.
authors: Yuki
authors: Y
authors: Ohmuro-Matsuyama
authors: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.
authors: Keita
authors: K
authors: Mori
authors: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.
authors: Hirotsugu
authors: H
authors: Hamada
authors: Chemical Resources Laboratory, Tokyo Institute of Technology, R1-18, 4259 Nagatsutacho, Midori-ku, Yokohama, 226-8503, Japan.
authors: Hiroshi
authors: H
authors: Ueda
authors: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan. yamaji@kobe-u.ac.jp.|Manufacturing Technology Association of Biologics (MAB), c/o Center for Business Incubation, Kobe University, 1-5-6 Minatojima-Minamimachi, Chuo-ku, Kobe, 650-0047, Japan. yamaji@kobe-u.ac.jp.
authors: Hideki
authors: H
authors: Yamaji
keywords: Antibody; Charged amino acid; Electrostatic steering; Fab fragment; Insect cell