鼠疫菌外膜蛋白单克隆抗体制备及初步应用

Prepation and application of monoclonal antibody to outer membrane of Yersinia pestis

应用ＳＤＳ－ＰＡＧＥ，蛋白免疫印迹（Ｗｅｓｔｅｒｎｂｌｏｔ）、单克隆抗体（ＭｃＡｂ）等技术，对鼠疫菌外膜蛋白（ＯＭＰ）组成成分及免疫学特性进行了研究、在ＳＤＳ－ＰＡＧＥ图谱上，鼠疫菌锡林郭勒高原型较其他型强、弱毒菌少一条亚基分子量为３２ＫＤ的蛋白带。以ＥＶ７６ｐ、ＥＶ苗的ＯＭＰ为免疫原，所获抗体进行Ｗｅｓｔｅｒｎｂｌｏｔ分析，认为ＯＭＰ具有良好的免疫原性及免疫反应性。采用杂交瘤技术，得到七株抗鼠疫菌的ＭｃＡｂ。四株为ＩｇＧ，三株为ＩｇＭ。经Ｗｅｓｔｅｒｎｂｌｏｔ鉴定。四株ＭｃＡｂ抗ＯＭＰ的分子量，４Ｇ５株为２１０ＫＤ，３Ｂ９、ＩＤ１０株为４０ＫＤ，２Ｃ６株为１０ＫＤ。以七株ＭＣＡｂ对鼠疫菌强、弱毒株３２株作全菌体ＥＬＩＳＡ进行鼠疫菌分型的初步研究发现，３Ｂ９株ＭｃＡｂ与２４株强毒菌发生阳性反应，而不与弱毒菌反应；１Ｄ１０株与锡型及弱毒菌不发生反应，与其它强毒菌发生阳性反应。

Using SDS-PAGE,Western blot and monoclonal antibody,the components and immunological features were studied of OMP.The OMP proved to a good antigen, with which its monoclonal antibody(McAb) was prepared and molecular weights of McAb were on demonstrated.The McAb was used in ELISA and a preliminary research was done on typing of Y.Pestis.