免疫亲和层析纯化汉坦病毒核壳蛋白实验条件的研究  

Study on the optimal dissociation conditions for immunoaffinity purification of nucleocapsid protein(NP)of Hantavirus(L99 strain)

作　　者：石晓宏[1] 宋干[1] 杭长寿[1] 林静 

机构地区：[1]中国预防医学科学院病毒学研究所,北京100052

出　　处：《中华实验和临床病毒学杂志》1996年第1期6-9,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：为了建立免疫亲和层析纯化汉坦病毒核壳蛋白(NP)的方法,应用酶联免疫吸附试验(ELISA)洗脱模型筛选出一种高效而温和的 NP 洗脱液,即 et-MgCl_2(乙二醇-MgCl_2)。et-MgCl_2能高效洗脱 NP,可在数分钟内将结合干 ELISA 板固相与 A35单克隆抗体(McAb)结合的 NP 几乎完全解离;在用 A35McAb-Sepharose 4B 免疫亲和层析柱纯化 NP 的实验中观察到同样的洗脱效果,经 SDS-PAGE 和 Western blot 证实,获得了高纯度的 L99株病毒 NP。In order to establish the optimal conditions for purifications of the structural proteins of Hantavirus by immunoaffinity chromatography,the efficient and non-denaturative eluents for NP were obtained by using ELISA elution model.It was found that the eluents for NP(et-MgCl_2)could dissociate NP quite efficiently within a few minutes,in which NP was combined with McAb A35 coated on ELISA plate.The elution efficiency was fur- ther confirmed by immunoaffinity purification of NP on A35-Sepharose 4B column.Highly purified and antigenically intact NP was recovered and identified by SDS-PAGE and Western Blot.

关 键 词：汉坦病毒 核壳蛋白 免疫亲和层析 

分 类 号：R446.5[医药卫生—诊断学]