免疫亲和层析提纯肾综合征出血热病毒蛋白

Purification of Viral Proteins of Hemorrhagic Fever with Renal Syndrome by Immunoaffinity Chromatography

本文应用免疫亲和层析从感染肾综合征出血热病毒(HFRSV)细胞中提纯了HFRSV蛋白。其最适条件:细胞混悬于磷酸盐缓冲生理盐水(PBS)—1％Tritonx—100、1％脱氧胆酸钠(DOC)、0.1％十二烷基硫酸钠(SDS)、1mM氟化磺酰苯甲烷(PMSF),超声波粉碎2分钟,5万r.p.m.1小时,离心上清液作为亲和层析的样品。亲和层析柱予先用PBS—1％Tritonx—100平衡,样品流经柱内亲和吸附后,再依次用PBS—1％Tritonx—100、PBS—0.1％Tritonx—100、PBS洗柱,最后用0.1M、pH11碳酸盐缓冲液洗脱。在上述条件下病毒蛋白得率高(91～97％),并保持其免疫活性。

The viral Proteins of hemorrhagic fever with renal syndrome were purified by immunoaffinity chromatography from virus-infected cells using the antibody of convalescent human sera from survivors of hemorrhagic fever with renal syndrome. Under optimal conditions the recovery of proteins of hemorrhagic fever virus reached 91—97.9％ and the pure proteins are immunologically competent.