Cell Reports Medicine (Jun 2021)
Highly specific monoclonal antibodies and epitope identification against SARS-CoV-2 nucleocapsid protein for antigen detection tests
- Yutaro Yamaoka,
- Kei Miyakawa,
- Sundararaj Stanleyraj Jeremiah,
- Rikako Funabashi,
- Koji Okudela,
- Sayaka Kikuchi,
- Junichi Katada,
- Atsuhiko Wada,
- Toshiki Takei,
- Mayuko Nishi,
- Kohei Shimizu,
- Hiroki Ozawa,
- Shuzo Usuku,
- Chiharu Kawakami,
- Nobuko Tanaka,
- Takeshi Morita,
- Hiroyuki Hayashi,
- Hideaki Mitsui,
- Keita Suzuki,
- Daisuke Aizawa,
- Yukihiro Yoshimura,
- Tomoyuki Miyazaki,
- Etsuko Yamazaki,
- Tadaki Suzuki,
- Hirokazu Kimura,
- Hideaki Shimizu,
- Nobuhiko Okabe,
- Hideki Hasegawa,
- Akihide Ryo
Affiliations
- Yutaro Yamaoka
- Department of Microbiology, Yokohama City University School of Medicine, Yokohama, Kanagawa 236-0004, Japan; Life Science Laboratory, Technology and Development Division, Kanto Chemical Co., Inc., Isehara, Kanagawa 259-1146, Japan
- Kei Miyakawa
- Department of Microbiology, Yokohama City University School of Medicine, Yokohama, Kanagawa 236-0004, Japan
- Sundararaj Stanleyraj Jeremiah
- Department of Microbiology, Yokohama City University School of Medicine, Yokohama, Kanagawa 236-0004, Japan
- Rikako Funabashi
- Department of Microbiology, Yokohama City University School of Medicine, Yokohama, Kanagawa 236-0004, Japan
- Koji Okudela
- Department of Pathology, Yokohama City University Graduate School of Medicine, Yokohama, Kanagawa 236-0004, Japan
- Sayaka Kikuchi
- Life Science Laboratory, Technology and Development Division, Kanto Chemical Co., Inc., Isehara, Kanagawa 259-1146, Japan
- Junichi Katada
- Medical Systems Research & Development Center, FUJIFILM Corporation, Kaisei, Kanagawa 258-8538, Japan
- Atsuhiko Wada
- Medical Systems Research & Development Center, FUJIFILM Corporation, Kaisei, Kanagawa 258-8538, Japan
- Toshiki Takei
- Medical Systems Research & Development Center, FUJIFILM Corporation, Kaisei, Kanagawa 258-8538, Japan
- Mayuko Nishi
- Department of Microbiology, Yokohama City University School of Medicine, Yokohama, Kanagawa 236-0004, Japan
- Kohei Shimizu
- Yokohama City Institute of Public Health, Yokohama, Kanagawa 236-0051, Japan
- Hiroki Ozawa
- Yokohama City Institute of Public Health, Yokohama, Kanagawa 236-0051, Japan
- Shuzo Usuku
- Yokohama City Institute of Public Health, Yokohama, Kanagawa 236-0051, Japan
- Chiharu Kawakami
- Yokohama City Institute of Public Health, Yokohama, Kanagawa 236-0051, Japan
- Nobuko Tanaka
- Yokohama City Institute of Public Health, Yokohama, Kanagawa 236-0051, Japan
- Takeshi Morita
- Department of Microbiology, Yokohama City University School of Medicine, Yokohama, Kanagawa 236-0004, Japan
- Hiroyuki Hayashi
- Division of Pathology, Yokohama Municipal Citizen’s Hospital, Yokohama, Kanagawa 221-0855, Japan
- Hideaki Mitsui
- Department of Pathology, Yokohama City University Graduate School of Medicine, Yokohama, Kanagawa 236-0004, Japan
- Keita Suzuki
- Life Science Laboratory, Technology and Development Division, Kanto Chemical Co., Inc., Isehara, Kanagawa 259-1146, Japan
- Daisuke Aizawa
- Life Science Laboratory, Technology and Development Division, Kanto Chemical Co., Inc., Isehara, Kanagawa 259-1146, Japan
- Yukihiro Yoshimura
- Division of Infectious Disease, Yokohama Municipal Citizen’s Hospital, Yokohama, Kanagawa 221-0855, Japan
- Tomoyuki Miyazaki
- Department of Physiology, Yokohama City University Graduate School of Medicine, Yokohama, Kanagawa 236-0004, Japan
- Etsuko Yamazaki
- Clinical Laboratory Department, Yokohama City University Hospital, Yokohama, Kanagawa 236-0004, Japan
- Tadaki Suzuki
- Department of Pathology, National Institute of Infectious Diseases, Shinjuku, Tokyo 162-8640, Japan
- Hirokazu Kimura
- School of Medical Technology, Faculty of Health Sciences, Gunma Paz University, Takasaki, Gunma 370-0006, Japan
- Hideaki Shimizu
- Division of Virology, Kawasaki City Institute for Public Health, Kawasaki, Kanagawa 210-0821, Japan
- Nobuhiko Okabe
- Division of Virology, Kawasaki City Institute for Public Health, Kawasaki, Kanagawa 210-0821, Japan
- Hideki Hasegawa
- Center for Influenza and Respiratory Virus Research, National Institute of Infectious Diseases, Musashimurayama, Tokyo 208-0011, Japan
- Akihide Ryo
- Department of Microbiology, Yokohama City University School of Medicine, Yokohama, Kanagawa 236-0004, Japan; Corresponding author
- Journal volume & issue
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Vol. 2,
no. 6
p. 100311
Abstract
Summary: The ongoing coronavirus disease 2019 (COVID-19) pandemic is a major global public health concern. Although rapid point-of-care testing for detecting viral antigen is important for management of the outbreak, the current antigen tests are less sensitive than nucleic acid testing. In our current study, we produce monoclonal antibodies (mAbs) that exclusively react with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and exhibit no cross-reactivity with other human coronaviruses, including SARS-CoV. Molecular modeling suggests that the mAbs bind to epitopes present on the exterior surface of the nucleocapsid, making them suitable for detecting SARS-CoV-2 in clinical samples. We further select the optimal pair of anti-SARS-CoV-2 nucleocapsid protein (NP) mAbs using ELISA and then use this mAb pair to develop immunochromatographic assay augmented with silver amplification technology. Our mAbs recognize the variants of concern (501Y.V1-V3) that are currently in circulation. Because of their high performance, the mAbs of this study can serve as good candidates for developing antigen detection kits for COVID-19.
