Construction and identification for cell strain of anti-human sperm protein 22 monoclonal antibodies.
- Author:
Xin-gang WANG
1
;
Chuan-dan WAN
;
Yu-feng HUANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; biosynthesis; immunology; Antibody Specificity; Blotting, Western; Cell Line; Enzyme-Linked Immunosorbent Assay; Female; Humans; Hybridomas; secretion; Immunohistochemistry; Male; Mice; Mice, Inbred BALB C; Microtubule-Associated Proteins; immunology; metabolism; Spermatozoa; metabolism
- From: National Journal of Andrology 2006;12(4):303-307
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare monoclonal antibodies of mice anti-human sperm protein 22 (SP22) and to identify their specificities.
METHODSBALB/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.
RESULTSThree 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.
CONCLUSIONThe 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.
