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Immunological detection of severe acute respiratory syndrome coronavirus by monoclonal antibodies
Authors:Ohnishi Kazuo  Sakaguchi Masahiro  Kaji Tomohiro  Akagawa Kiyoko  Taniyama Tadayoshi  Kasai Masataka  Tsunetsugu-Yokota Yasuko  Oshima Masamichi  Yamamoto Kiichi  Takasuka Naomi  Hashimoto Shu-ichi  Ato Manabu  Fujii Hideki  Takahashi Yoshimasa  Morikawa Shigeru  Ishii Koji  Sata Tetsutaro  Takagi Hirotaka  Itamura Shigeyuki  Odagiri Takato  Miyamura Tatsuo  Kurane Ichiro  Tashiro Masato  Kurata Takeshi  Yoshikura Hiroshi  Takemori Toshitada
Affiliation:Department of Immunology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
Abstract:In order to establish immunological detection methods for severe acute respiratory syndrome coronavirus (SARS-CoV), we established monoclonal antibodies directed against structural components of the virus. B cell hybridomas were generated from mice that were hyper-immunized with inactivated SARS-CoV virion. By screening 2,880 generated hybridomas, we established three hybridoma clones that secreted antibodies specific for nucleocapsid protein (N) and 27 clones that secreted antibodies specific for spike protein (S). Among these, four S-protein specific antibodies had in vitro neutralization activity against SARS-CoV infection. These monoclonal antibodies enabled the immunological detection of SARS-CoV by immunofluorescence staining, Western blot or immunohistology. Furthermore, a combination of monoclonal antibodies with different specificities allowed the establishment of a highly sensitive antigen-capture sandwich ELISA system. These monoclonal antibodies would be a useful tool for rapid and specific diagnosis of SARS and also for possible antibody-based treatment of the disease.
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