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目的制备用于检验人精浆特异蛋白P30—主要是来自法医学案件的免疫胶体金层析试剂条。方法选取针对不同抗原决定簇的抗P30单克隆抗体细胞株,并制备其小鼠腹水,分离纯化单克隆抗体。制备胶体金并以纯化抗体包被,制成免疫胶体金,以免疫胶体金浸泡玻璃纤维。选取适宜的硝酸纤维素膜并于其上不同位置以未金标的另一株P30单克隆抗体和羊抗鼠IgG包被。搭建试剂条并检测其灵敏度和特异性。结果所制成的试剂条灵敏度至少可达4ng/ml;对6人份混合的人精液物质在稀释20万倍后仍出阳性结果,且无非特异性反应。结论检验人精浆特异蛋白P30的免疫胶体金试剂条可对嫌疑人精物质做出排查,有利于法医物证检验。
Objective To prepare an immuno-colloidal gold chromatography reagent strip for testing human serum-specific protein P30 - mainly from forensic medical cases. Methods Anti-P30 McAbs against different antigenic determinants were selected and their mouse ascites were prepared. The monoclonal antibodies were isolated and purified. Preparation of colloidal gold and purified antibody coated, made of immune colloidal gold, immune colloidal gold soaked glass fiber. A suitable nitrocellulose membrane was selected and coated at another location on top of it with another gold-free P30 monoclonal antibody and goat anti-mouse IgG. Set up a reagent strip and test its sensitivity and specificity. Results The sensitivity of the prepared strips was at least 4ng / ml; for human semen mixed with 6 samples, there was still positive results after diluted 200000 times and there was no nonspecific reaction. Conclusion The test of the human serum-specific protein P30 immuno-colloidal gold reagent strips can make investigation of the suspicious substances, which is in favor of forensic evidence.