title: Good manufacturing practices production of a purification-free oral cholera vaccine expressed in transgenic rice plants.
abstract: The first Good Manufacturing Practices production of a purification-free rice-based oral cholera vaccine (MucoRice-CTB) from transgenic plants in a closed cultivation system yielded a product meeting regulatory requirements. Despite our knowledge of their advantages, plant-based vaccines remain unavailable for human use in both developing and industrialized countries. A leading, practical obstacle to their widespread use is producing plant-based vaccines that meet governmental regulatory requirements. Here, we report the first production according to current Good Manufacturing Practices of a rice-based vaccine, the cholera vaccine MucoRice-CTB, at an academic institution. To this end, we established specifications and methods for the master seed bank (MSB) of MucoRice-CTB, which was previously generated as a selection-marker-free line, evaluated its propagation, and given that the stored seeds must be renewed periodically. The production of MucoRice-CTB incorporated a closed hydroponic system for cultivating the transgenic plants, to minimize variations in expression and quality during vaccine manufacture. This type of molecular farming factory can be operated year-round, generating three harvests annually, and is cost- and production-effective. Rice was polished to a ratio of 95 % and then powdered to produce the MucoRice-CTB drug substance, and the identity, potency, and safety of the MucoRice-CTB product met pre-established release requirements. The formulation of MucoRice-CTB made by fine-powdering of drug substance and packaged in an aluminum pouch is being evaluated in a physician-initiated phase I study.
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.|Engineering Headquarters, Asahi Kogyosha Co., Ltd., 3-13-12, Mita, Minato-ku, Tokyo, 108-0073, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Kashima
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan. yukiy@ims.u-tokyo.ac.jp.|International Research and Development Center for Mucosal Vaccine, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan. yukiy@ims.u-tokyo.ac.jp.
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Yuki
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.|Seto Center, Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, 4-1-70, Seto-Cho, Kanonji, Kagawa, 768-0065, Japan.
authors.forename: Mio
authors.initials: M
authors.lastname: Mejima
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.|Seto Center, Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, 4-1-70, Seto-Cho, Kanonji, Kagawa, 768-0065, Japan.
authors.forename: Shiho
authors.initials: S
authors.lastname: Kurokawa
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.|Seto Center, Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, 4-1-70, Seto-Cho, Kanonji, Kagawa, 768-0065, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Suzuki
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.|Seto Center, Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, 4-1-70, Seto-Cho, Kanonji, Kagawa, 768-0065, Japan.
authors.forename: Satomi
authors.initials: S
authors.lastname: Minakawa
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.|Research Department, Nippon Institute for Biological Science, 9-2221-1, Shin-machi, Ome, Tokyo, 198-0024, Japan.
authors.forename: Natsumi
authors.initials: N
authors.lastname: Takeyama
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Fukuyama
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Azegami
authors.affiliation: Seto Center, Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, 4-1-70, Seto-Cho, Kanonji, Kagawa, 768-0065, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tanimoto
authors.affiliation: Crop Development Division, NARO Agriculture Research Center, 1-2-1, Inada, Joetsu-shi, Niigata, 943-0193, Japan.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Kuroda
authors.affiliation: Engineering Headquarters, Asahi Kogyosha Co., Ltd., 3-13-12, Mita, Minato-ku, Tokyo, 108-0073, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Tamura
authors.affiliation: Seto Center, Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, 4-1-70, Seto-Cho, Kanonji, Kagawa, 768-0065, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Gomi
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.|International Research and Development Center for Mucosal Vaccine, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kiyono
mesh_terms: D000284:Administration, Oral; D000818:Animals; D015153:Blotting, Western; D002771:Cholera / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control; D002772:Cholera Toxin / Q000633:toxicity; D022121:Cholera Vaccines / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology; D003362:Cost-Benefit Analysis; D003967:Diarrhea / Q000139:chemically induced / Q000276:immunology / Q000517:prevention & control; D004349:Drug Packaging; D004355:Drug Stability; D006801:Humans; D007114:Immunization / Q000379:methods; D051379:Mice; D012275:Oryza / Q000235:genetics* / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000254:growth & development; D011208:Powders; D015203:Reproducibility of Results; D013678:Technology, Pharmaceutical / Q000191:economics / Q000379:methods*; D014734:Vibrio cholerae / Q000276:immunology
chemical_list: D022121:Cholera Vaccines; D011208:Powders; D002772:Cholera Toxin
keywords: GMP production; Molecular farming; MucoRice; Oral vaccine; Seed bank