Document Detail

The sperm acrosome: immunological analysis using specific polyclonal and monoclonal antibodies directed against the outer acrosomal membrane of boar spermatozoa.
MedLine Citation:
PMID:  3526982     Owner:  NLM     Status:  MEDLINE    
An antiserum to the purified porcine outer acrosomal membrane (OAM) was raised in female Balb/c mice and was characterized by means of an indirect ELISA. The hyperimmune serum reacted selectively with the acrosomal cap of the sperm head and showed an extremely good cross reactivity with bull and human spermatozoa when assayed by indirect immunofluorescence. Immunoelectron microscopy using the protein A-gold method further confirmed the specificity of the anti-OAM-antiserum for the OAM. In an effort to identify the OAM antigens recognized by the hyperimmune serum and to analyse the extent of cross reactivity on a molecular level, the SDS-extractable proteins were separated by SDS-PAGE, transblotted and immunoprinted using an 125J-conjugated anti-mouse-antibody. To facilitate functional and structural analysis of distinct OAM-proteins monoclonal antibodies were generated by hybridization of mouse myeloma cells with the splenocytes of female Balb/c mice immunized with the purified OAM. One fusion resulted in about 100 anti-OAM-antibodies secreting hybridoma cultures, of which about 30% showed cross reaction with human and bull spermatozoa. Four stable cell lines were selected for this study secreting antibodies directed against the outer acrosomal membrane of boar spermatozoa. Whereas the polyclonal immune mouse serum stained the entire acrosomal cap, the four hybridoma antibodies generated a patch-work-like immunofluorescence pattern over the acrosome. HPLC-ELISA of the solubilized OAM revealed first information on the nature of the corresponding membrane antigen.
E Töpfer-Petersen; J Auerbeck; A Weiss; A E Friess; W B Schill
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Andrologia     Volume:  18     ISSN:  0303-4569     ISO Abbreviation:  Andrologia     Publication Date:    1986 May-Jun
Date Detail:
Created Date:  1986-09-17     Completed Date:  1986-09-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0423506     Medline TA:  Andrologia     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  237-51     Citation Subset:  IM    
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MeSH Terms
Acrosome / immunology*,  ultrastructure
Antibodies / immunology*
Antibodies, Monoclonal / immunology*
Cell Membrane / immunology
Chromatography, High Pressure Liquid
Enzyme-Linked Immunosorbent Assay
Fluorescent Antibody Technique
Mice, Inbred BALB C / immunology
Spermatozoa / immunology*
Reg. No./Substance:
0/Antibodies; 0/Antibodies, Monoclonal

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