蛇毒C型凝集素类似蛋白Agkisacutacin单克隆抗体的制备和鉴定

Preparation and identification of monoclonal antibodies against snake venom C-type lectin like protein Agkisacutacin

目的:制备并鉴定抗蛇毒C型凝聚素类蛋白Agkisacutacin的单克隆抗体(mAb)。方法:以Agkisacutacin蛋白为抗原免疫BALB/c小鼠,通过常规的细胞融合方法制备抗Agkisacutacin蛋白的mAb。用间接ELISA法检测杂交瘤细胞分泌mAb的稳定性。用荧光染色法做杂交瘤细胞的核型分析。用Western blot检测mAb的特异性。结果:获得3株可稳定分泌抗AgkisacutacinmAb的杂交瘤细胞株。结论:成功制备了抗Agkisacutacin的mAb,为检测Agkisacutacin蛋白作为新型抗血栓药物的体内代谢规律的方法和研究抗血栓的机制提供了重要的工具。

AIM： To prepare and identify the antibodies against snake venom C-type lectin like protein Agkisacutacin. METHODS： A BALB/c mouse was immunized with Agkisacutacin and the cells were fused by standard hybridoma technique to prepare mAbs. The stability of the hybridomas secreting mAbs was detected by indirect ELISA. The nuclear type of the hybddomas was analyzed by fluorescent staining and the specificity of mAbs was detected by Westem blot. RESULTS. Three cell strains of hybridomas that could steadily secrete anti-Agkisacutacin mAb were obtained. CONCLUSION： The successful preparation of antiAgkisacutacin mAbs provides an important tool for studying the in vivo metabolism of new anti-thrombus drugs by detecting Agkisacutacin and investigating the mechanism of antihrombus of the drugs.