流行性出血热病毒核蛋白的血凝位点研究

The Study of Hemagglutination Epitopes on Nucleocapsid Protein of Epidemic Hemorrhagic Fever Virus

流行性出血热病毒（ＥＨＦＶ）Ａ９株鼠脑抗原，经Ａ３５ＭｃＡｂ偶联于Ｓｅｐｈａｒｏｓｅ４Ｂ制备的免疫吸附柱进行亲和层析纯化，并用高压液相色谱（ＨＰＬＣ）进一步纯化，获得了高纯度的病毒核衣壳蛋白（ＮＰ），其分子量为５００００，经不同特性的ＭｃＡｂ和ＥＨＦ患者血清通过间接ＥＬＩＳＡ和Ｗｅｓｔｅｒｎ－ｂｌｏｔ鉴定，证实为ＥＨＦ病毒特异性的ＮＰ，且具有血凝活性。将ＨＰＬＣ纯化的ＮＰ免疫小鼠后，可诱导产生特异性血凝抑制抗体，但无中和抗体产生，由此证实ＥＨＦ病毒（Ａ９株）ＮＰ具有构型依赖性血凝活性结合位点。

Epidemic Hemorrhagic Fever virus (A9 strain) antigen was purified by affinity chromatography prepared with A35 McAb against nucleocapsid(NP) of EHF virus linking to sepharose 4B and further isolated and purified by high pressure liquid chromatography (HPLC).The antigen purified by HPLC had higher purity and was determined to be EHF vrius specific NP with many McAbs and EHF patient sera by indirect-ELISA and western blot, M. W. of which was 50kD. HPLC-NP had still hemagglutination activity. The hemagglutination-inhibiting antibody could be detected in the immune sera with HPLC-NP,which proved that there are conformation depending hemagglutination epitopes.