[Preparation and characterization of monoclonal antibody against Vibrio cholerae O139].
OBJECTIVE
To prepare monoclonal antibody (mAb) against Vibrio cholerae O139, which would be used as the gold colloidal reagent strip for rapid detection of O139, and to determine its biological characterization.


METHODS
BALB/c mice were immunized by inactivated Vibrio cholerae O139. Anti-O139 mAbs were prepared by using hybridoma technique. The specificity of mAb was determined by indirect ELISA and Western blot. The indirect ELISA was used to identify Ig subgroup, detect its titer in ascites and relative affinity, and analyze antigen-binding epitope of mAbs.


RESULTS
Two hybridoma cells (O4D7 and O4D10), secreting anti-O139 mAbs were obtained. The Ig subgroups of O4D7 and O4D10 were IgG2b and IgG3, respectively. The mAb's titer in ascites was 1:10(7). The relative affinity of mAb O4D7 was more than 10(5) and that of O4D10 was more than 10(4). The result of additive ELISA showed that two mAbs could recognize different antigen epitopes.


CONCLUSIONS
The successful preparation of two anti-O139 mAbs provides a powerful tool for the development of a rapid method of detecting Vibrio cholerae O139 infection.
Zhang
Jun
J
Key Laboratory of Infectious Diseases of the Ministry of Education, Chongqing Medical University, Chongqing 400016, China. zhangjunlg@hotmail.com
Zhang
Juan
J
Tang
Ni
N
Zhang
Bing-qiang
BQ
Chen
Wei-xian
WX
Huang
Ai-long
AL
Zheng
Jian
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000917:Antibody Formation / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D008807:Mice, Inbred BALB C; D015164:Vaccines, Inactivated / Q000008:administration & dosage* / Q000276:immunology; D041242:Vibrio cholerae O139 / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015164:Vaccines, Inactivated