[Preparation, characterization and preliminary application of monoclonal antibodies against the expressed Norovirus capsid protein].
OBJECTIVE
To prepare the monoclonal antibodies (mAbs) against Norovirus capsid protein for the development of a rapid assay of Norovirus and to investigate the pathogenesis of this virus.


METHODS
Sp2/0-Ag-14 myeloma cells were fused with spleen cells of BALB/c mice immunized with the recombinant protein of Norovirus NVgz01 (DQ369797), which was overexpressed in E.coli. The monoclonal antibodies against Norovirus capsid protein were screened by selective culture medium. The obtained Ig subtypes, titer and specificity of the mAbs were examined by ELISA and Western blot respectively.


RESULTS
After cell fusion and subcloning, four hybridoma cell lines which secreted monoclonal antibodies specifically against Norovirus capsid protein were obtained and named as N2C3, N7C2, N4B1 and N8A9. Indirect ELISA and Western blot assay showed that the four mAbs specifically recognized the Norovirus capsid protein expressed in E.coli. The native Norovirus capsid protein in the stool samples were also recognized by them. The Sandwich ELISA, a rapid detection assay of the expressed and native Norovirus capsid protein, demonstrated that N2C3 and N7C2 were matched successfully.


CONCLUSIONS
The mAbs against GII Norovirus capsid protein have been developed, which can be used for the development of detection assay and the basic research of Norovirus.
Li
Xiao
X
South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China. xiao-2001@21cn.com
Zhou
Rong
R
Sheng
Hui-Ying
HY
Wang
Chang-Bing
CB
Tian
Xin-Gui
XG
Wang
You-Shao
YS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D008807:Mice, Inbred BALB C; D029322:Norovirus / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D036022:Capsid Proteins