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  • Title: Crossed immunoelectrophoresis of sperm antibodies in human serum and cervical mucus.
    Author: Gradl T, Mettler L.
    Journal: Int J Fertil; 1975; 20(1):50-4. PubMed ID: 4387.
    Abstract:
    Sperm agglutinating antibodies are purified from sera and cervical mucus of women with unexplained causes of infertility which were positive in the FD-test. Fractionation was performed by affinity-chromatography in a batch device and the sperm agglutinating activity controlled by the Franklin and Dukes test. This sperm antibody fraction was determined via crossed immunoelectrophoresis by migration into an anti-human serum containing gel. In all cases only one big peak resulted. The negative control serum and mucus samples demonstrated no precipitation peaks. By absorption studies it was shown that the sperm agglutinating antibodies in sera were IgM and in cervical mucus IgA. The concentration of IgA and IgM was determined by comparison with standard human IgA and IgM. Thus only one serum- and one cervical mucus antibody seems to be responsible for agglutination. The number of experiments, however, is still too small for general conclusions. This method is easily and quickly performed and can therefore be used as a routine method for the determination of sperm agglutinating antibodies. Its application for sperm-immobilizing or cytotoxic activity remains to be tested. A new technique by which sperm antibodies are purified by affinity-chromatography in a batch device and the sperm agglutinating activity controlled by the Franklin and Dukes (FD) test is described. Specific absorption takes place on an insoluble antigen and desorption reveals purified antibodies. In sera of women with unexplained infertility which were found to be positive to the FD test a specific globulin binding takes place. For quantitative detection of these purified antibodies, crossed immunoelectrophoresis was done. Sera of 4 such infertile women were used. Uterine cervical mucus samples of 3 of them were also examined. As controls, sera negative to the FD test and cervical mucus from the same patients were processed. Spermatozoa were obtained from 15 donors, pooled, washed 3 times with phosphate-buffered saline, and adjusted to spermatozoa content. Cervical mucus was also prepared. Techniques used are given. The method can be easily and quickly performed and therefore used routinely. Antibody fractions, purified by affinity-chromatography migrating against an AHS-containing gel, demonstrated 1 big peak in all 4 of the FD-positive semen samples as well as in the 3 cervical mucus samples. The height of the peak represented a nearly linear relation to the sperm antibody content of the semen. Control samples never showed this peak. By absorption experiments it was found that the protein encountered in serum is IgM and that in cervical mucus IgA. The content of antibodies in the cervical mucus of the same woman was about 1/10 smaller than in her serum. This method is stated to be an improvement over other techniques. Some sperm antibodies are not of the agglutinating type.
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