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Updated: Aug 8, 2026
Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
Published on: March 24, 2017
[Construction and identification for cell strain of anti-human sperm protein 22 monoclonal antibodies]
Xin-gang Wang1, Chuan-dan Wan, Yu-feng Huang
1Department of Microbiology and Immunology, NJMU, Nanjing, Jiangsu, China.
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.
Conclusion:
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.
