Preparation and identification of anti-human epididymal sperm protein P34H monoclonal antibody / 中华男科学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To prepare and identify anti-human epididymal sperm protein P34H monoclonal antibody (McAb).</p><p><b>METHODS</b>The previously purified recombinant P34H protein was used as an antigen to immunize BALB/c mouse. Cell strains secreting anti-P34H McAb were established by hybridoma technique and then ascitic fluid-type McAb prepared. The sensitivity and specificity of McAb were detected by ELISA and Western blot, respectively.</p><p><b>RESULTS</b>One strain (2C4) of IgG1 Kappa anti-P34H McAb was harvested. The titer detected by ELISA technique was 110(3) - 110(5). Western blot of healthy human sperm samples and purified recombinant P34H antigen probed with the prepared McAb were immunoreactive at 34,000 and 27,000, respectively.</p><p><b>CONCLUSION</b>Anti-P34H McAb has been prepared successfully by the above methods, which may provide a powerful tool for exploring the relationship between P34H and male fertility.</p>