蛇毒降纤酶多克隆抗体的制备及其某些性质

Preparation of polyclonal antibody against defibrase from Agkistrodon halys ussurriensis Emelianov snake venom

为制备长白山白眉蝮蛇蛇毒降纤酶的多克隆抗体 ,以期获得酶免疫测定所需的反应试剂 ,供药剂学的研究应用。经免疫兔获得高效价的抗长白山白眉蝮蛇蛇毒降纤酶的抗血清 ,以环状沉淀反应、琼脂免疫双扩散 ,以及酶联免疫吸附法 ,验证了产生的抗体。免疫双扩散及ELISA测定的结果表明 ,长白山白眉蝮蛇蛇毒降纤酶与日本克栓酶有着不同的抗原决定簇结构。 3只兔的抗血清以 1∶7 2万的倍数稀释时 ,其在ELISA测定中的光密度 (OD4 90nm)仍比空白对照高出 0 2以上 ,具有较高效价 ;该抗体与精制蝮蛇抗栓酶有一定的交叉反应 (比率为 0 0 2 4) ,与江浙蝮蛇蛇毒粗品及日本克栓酶间的交叉反应微小 (比率小于 0 0 0 1) 。

Polyclonal antibodies to Ahue def were developed in three rabbits. The results of capilary immunoprecipitation, double immunodiffusion and ELISA, showed the generation of high titer rabbit antibodies against Ahue def. The antibodies showed high specifity to Ahue def, exhibiting slight cross reactivity with the purified kangshuanmei (ksm) and only very limited cross reactivity with the Japanese Defibrase or Jiangzhe crude snake venom. The results also implied that differences in the immunological structure existed between Ahue def from agkistrodon halys ussurriensis emilianov venom and the Japanese Defibrase.