[Construction and identification for cell strain of anti-human sperm protein 22 monoclonal antibodies].
OBJECTIVE
To prepare monoclonal antibodies of mice anti-human sperm protein 22 (SP22) and to identify their specificities.


METHODS
BALB/c mice were immunized with human SP22, monoclonal antibodies were prepared by hybridoma technique and the sensitivity and specificity of SP22 McAb were investigated by ELISA and Western-blot assay, respectively. The distribution of human SP22 in sperm were shown by immunohistochemistry.


RESULTS
Three strains of hybridoma cells were obtained, with the affinity constant (K) of 1.0 x 10(7) L/mol and the titers were 1:10(3) and 1:3,200 in the mixed supernatant of cell cultures and abdominal dropsy, respectively. IgG isotype of the antibody was identified as IgG1. Western blot demonstrated that there was a specific recognition between human SP22 and the obtained monoclonal antibody. Immunohistochemistry displayed that human SP22 mainly distributed on the acrosome surface of human sperm.


CONCLUSIONS
The monoclonal antibodies of anti-human 22 was prepared by the technique of hybridoma cell has higher titer and specificity, which can combine specially with the SP22 protein on the surface of human sperm.
Wang
Xin-gang
XG
Department of Microbiology and Immunology, NJMU, Nanjing, Jiangsu, China.
Wan
Chuan-dan
CD
Huang
Yu-feng
YF