单克隆抗体亲和层析法纯化HFRS病毒血凝素抗原的初步研究

用蔗糖-丙酮法从HFRS病毒陈栋感染的乳小白鼠鼠脑中提取粗制血凝素,将此粗制血凝素通过抗HFRS病毒血凝素McAb 3G_1与Sepharose 4B偶联的免疫吸附柱,获得初步纯化的HFRS病毒血凝素抗原,经鉴定具有较高的血凝滴度和较好的免疫原性。用SDS-PAGE和Wes-tern-blot技木分析该纯化的血凝素抗原,并与同法纯化的正常鼠脑抗原比较,证明该特异性血凝素抗原分子量为5×10~4道尔顿。用2株抗HFRS病毒以及10株抗HFRS病毒血凝素McAb,以ELISA阻断试验分析纯化50K血凝素,结果有10株McAb可阻断McAb3 G_1与其结合,其抗原决定簇之间有部分相同或重叠,提示这些具有不同特性的抗原决定簇确实位于同一结构蛋白上。以上结果表明,该50k血凝素蛋白特性及结构均较复杂,尚待进一步研究。

Purification of Hemagglutinin Antigen of the HFRS Virus by Affinity ChromatograPhy with McAb

HFRS virus hemagglutinin was harvested from suckling mice brain inoculated with HPKS virus Chen's strain and prepared in high titre hemagglutinin antigen by sucrose-acetone extraction method. The roughly purified hemagglutinin was then passed to immu-noadsorbent eoluram prepared by sepharose-IB coupled with McAb 3Gl against HFRS virus hemagglutinin.Wheu the column was eluted with potassium thiocyanate a specifical hemagglutinin antigen of HFRS virus was harvested and the eluted fraction were identified as specifical hcmagglulinin antigen by the specifical antigeu-blocking ELISA and the replacing test.The hemagglutinin antigen was piovidcd with high titer and good immuno-genicity. The purificatory hemagglutinin was analysed and compare with normal mouse brain antigen by SDS-PAGE and the Western-blotting test using HRP-McAb 3Gl immuno-cnzymc technic and its molecular weight being 50 000 dalton.The 50K hemagglutinin antigen was analysed by an ELISA blokiug test using 12 McAbs specific for HFRS virus and gcmagglutinin. It was showed that 10 McAbs could blocked reaction of McAb 3Gl with the solv hemagglutinin. It was proved that there are differences among the antigenic the 50K proiein directed by McAbs were partly identical or overlapping and were located on same a polypiptide chain. Analysis of the results shows that the characteristics and structure of the 50K heuiagglutiniu antigen are complex and need go deep into study.