尖吻蝮蛇出血毒素Ⅲ分离、抗血清制备、与其他出血毒素血清学关系研究

Agkistrodon acutus Venom,Antiserum Preparation and Serological Relationships among other Haemorrhagins in Agkistrodon acutus Venom

先用DEAE-Sephadex A 50离子交换层析,然后用CM-Sephadex C 50离子交换层析,最后用12%SDS-PAGE制备电泳纯化尖吻蝮蛇出血毒素Ⅲ。将含出血毒素Ⅲ的聚丙烯酰胺凝胶磨细免疫小鼠,制备抗血清。ELISA检测显示制备的抗血清IgG能够与天然的出血毒素Ⅲ结合,Western blot检测结果显示制备的抗血清只与出血毒素Ⅲ特异结合,制备的抗血清可以用于免疫组织化学实验,为以后检测血毒素Ⅲ的结合位点打下了基础。

Objective：Antiserum against haemorrhaginⅢfromAgkistrodon acutus venom was prepared for studying serological relationships among other haemorrhagins in Agkistrodon acutus venom and detecting haemorrhaginⅢ binding site in animal body in future.Methods：HaemorrhaginⅢ was purified firstly by DEAE-Sephadex A 50,subsequently by CM-Sephadex C 50,and further purified by 12%SDS-PAGE.The antiserum against haemorrhaginⅢ was raised in mouse and its specificity was confirmed by Western blot analysis.The ability to combine with nature haemorrhaginⅢ was confirmed by ELISA analysis.Results：The antiserum is special to haemorrhagin III and IgG could combine haemorrhaginⅢ.Conclusion：The antiserum prepared in this study is suitable for haemorrhagin III detection.