Immunolocation of antisperm monoclonal antibody 6B10 and corresponding antigen |
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Authors: | GAO Shaorong HU Guojun DUAN Chongweni LIU Hui HAN Zhiming SONG Xiangfen CHEN Dayuan |
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Institution: | State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100080, Beijing, China. |
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Abstract: | An antisperm monoclonal antibody 6B10 was produced by hybridoma technique of the isotype IgG. The monoclonal antibody was purified by means of ammonium sulfate precipitation and protein A-Sepharose C1-4B affinity chromatography. SDS polyacrylamide gel electrophoresis was used to evaluate the purity of the antibody. Evaluation of the sperm acrosomal status was determined by chlortetracycline (CTC) staining. It was found that monoclonal antibody 6B10 can inhibit the sperm acrosome reaction induced by progesterone. The corresponding antigen recognized by monoclonal antibody 6B10 was located on the plasma membrane of the sperm acrosome by indirect immunofluorescent microscopy and immunoelectronmicroscopy. Sperm protein was extracted by 1 % Triton X-100. The molecular weight of the antigen is 50 ku, detected by Western blot. The antigen is a key protein in the sperm acrosome reaction and may be the receptor of progesterone on the sperm acrosome. It may either be developed as a candidate contraceptive vaccine or be used as a tool in pest/rodent management. |
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Keywords: | monoclonal antibody hybridoma technique progesterone antigen acrosome reaction |
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