Immunological detection of severe acute respiratory syndrome coronavirus by monoclonal antibodies.
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.
Ohnishi
Kazuo
K
Department of Immunology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
Sakaguchi
Masahiro
M
Kaji
Tomohiro
T
Akagawa
Kiyoko
K
Taniyama
Tadayoshi
T
Kasai
Masataka
M
Tsunetsugu-Yokota
Yasuko
Y
Oshima
Masamichi
M
Yamamoto
Kiichi
K
Takasuka
Naomi
N
Hashimoto
Shu-ichi
S
Ato
Manabu
M
Fujii
Hideki
H
Takahashi
Yoshimasa
Y
Morikawa
Shigeru
S
Ishii
Koji
K
Sata
Tetsutaro
T
Takagi
Hirotaka
H
Itamura
Shigeyuki
S
Odagiri
Takato
T
Miyamura
Tatsuo
T
Kurane
Ichiro
I
Tashiro
Masato
M
Kurata
Takeshi
T
Yoshikura
Hiroshi
H
Takemori
Toshitada
T
D000818:Animals; D000911:Antibodies, Monoclonal; D002522:Chlorocebus aethiops; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis*; D014709:Vero Cells
D000911:Antibodies, Monoclonal