以原核细胞表达的人PrP蛋白为抗原制备抗人朊蛋白抗体  

Preparation of polyclonal antibody to human prion protein using the expressed GST-PrP fusion protein as antigen

作　　者：赵霞[1] 董小平[1] 周伟[1] 赵同兴[1] 洪涛[1] 

机构地区：[1]中国预防医学科学院病毒学研究所形态室,北京100052

出　　处：《中华实验和临床病毒学杂志》2000年第2期131-133,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家 8 63高科技发展计划资助项目!( 863 10 2 11 0 3 0 5 )

摘　　要：目的　制备针对朊蛋白的特异性抗体。方法　以初步纯化的原核细胞表达的GST PrP融合蛋白免疫家兔 ,制备朊蛋白特异的抗体。结果　以GST PrP s为抗原 ,用酶联免疫吸咐试验(ELISA)检测免疫家兔血清中的朊病毒抗体效价最高可达至 12 80 0 0。Westernbolt检测显示所制备的抗体不仅可以与体外表达的全长及不同C 末端缺失的人朊蛋白反应 ,而且能够识别人、鼠脑组织中内源性朊蛋白。结论　利用GST PrP融合蛋白可有效地剌激免疫动物产生PrP特异性抗体 ,所制备的抗血清可试用于朊蛋白的检测。Objective Preparing specific antibody to prion protein.Method Using prokaryotic expressed GST PrP fusion protein as antigen, rabbits were immunized subcutaneously.Results ELISA assay revealed that the titer of the prepared antiserum against expressed PrP protein was as high as 1:128 000. Western blot test showed that the antiserum was able to react with the in vitro expressed intact and different lengths of C terminus truncated PrP proteins, as well as the native PrP proteins from brain homogenization of human and mouse.Conclusion The prokaryotic expressed GST PrP fusion protein can efficiently elicit in immunized animals the PrP specific antibody.

关 键 词：朊病毒 病毒蛋白质 病毒抗体 

分 类 号：R392-33[医药卫生—免疫学] R373.9[医药卫生—基础医学]