Anti-norovirus polyclonal antibody and its potential for development of an antigen-ELISA.
Norovirus (NoV) capsid proteins were expressed as virus-like particles (VLPs) by using recombinant baculovirus in insect cells, which had 5 genotypes in genogroup I and 11 genotypes in genogroup II, and the VLPs were used as immunogens. Polyclonal antibody against the VLP of GII/3 genotype showed broad-range cross-reactivity, reacting not only with intra-genogroup strains, but also inter-genogroup strains, by antibody-ELISA using 16 kinds of VLPs. Furthermore, antigen-ELISA was conducted in sandwich enzyme-linked immunosorbent assay (ELISA) using the polyclonal antibody for capturing antigens, and three kinds of monoclonal antibodies against the VLP of GII/4 genotype for detecting antigens. This format successfully detected eight genotypes of NoV from clinical specimens and proved that polyclonal antibody, which has broad-range cross-reactivity, was capable of detecting various types of genotypes from clinical specimens.
Okame
Michio
M
Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Shiota
Tomoyuki
T
Hansman
Grant
G
Takagi
Makiko
M
Yagyu
Fumihiro
F
Takanashi
Sayaka
S
Phan
Tung Gia
TG
Shimizu
Yuko
Y
Kohno
Hideki
H
Okitsu
Shoko
S
Ushijima
Hiroshi
H
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005243:Feces / Q000821:virology; D005838:Genotype; D006801:Humans; D029322:Norovirus / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral