A SOLID-PHASE ASSAY FOR THE DETECTION OF ANTISPERM ANTIBODIES

PROBLEM: ELISA is an ideal assay method for a large-scale screening of anti-sperm antibodies among a large number of infertile males. Howeve r, conventional ELISA with whole spermatozoa needs time-consuming step s of centrifugation. METHOD: A solid-phase assay used for detecting an ti-sperm antibodies was established. This assay is suitable not only f or detecting circulating anti-sperm antibodies of IgG, IgM, and IgA su bclass simultaneously but also for screening hybridomas secreting anti -sperm monoclonal antibodies (mAbs). The microtiter plates, on which s olubilized sperm antigens are fixed, can be stored at -80-degrees-C fo r up to six months without losing reactivity with anti-sperm antibodie s. RESULTS: Using this assay, 53 sera (13 were proven positive and 40 were proven negative for sperm agglutination antibody) were tested. Al though the false-negative rate was 0%, the false-positive rate was 32% . One thousand one hundred sixty-five supernatants from hybridomas con structed with splenocytes of mice who were hyperimmunized with human s perm and nonsecreting myeloma cells were tested by this solid-phase as say and two anti-sperm mAb secreting clones were selected and establis hed. CONCLUSIONS: It is recommended that for research work this assay could be used for the first screening of the hybridoma secreting anti- sperm mAb, and for clinical use this assay might be suitable for the f irst screening of sera of infertile patients. However, conventional bi oassays should follow to confirm the biological meaning of the positiv ity.