汉坦病毒G2糖蛋白多表位抗原的表达及应用

Expression and use of a multi-epitope antigen derived from the G2glycoprotein of Hantavirus

目的对汉坦病毒G2糖蛋白多表位抗原(multi-epitope antigen,MEA)进行原核表达和纯化,建立MEAELISA方法并用于肾综合征出血热(HFRS)患者血清特异性抗体的检测。方法利用pET原核表达系统表达MEA,并进行镍亲和纯化;利用Western blot方法对MEA进行免疫反应性鉴定,并建立MEA-ELISA方法,用于检测HFRS患者血清特异性抗体。结果原核表达系统E.coli BL21/pET32a-mea能表达分子质量单位为36.9ku的MEA蛋白,纯化后的蛋白浓度为0.15mg/ml,纯度>95%,Western blot显示目的蛋白能被HFRS患者血清识别。用建立的MEA-ELISA方法检测120例HFRS患者血清的阳性率为90.83%,检测60例健康人血清全部为阴性。结论重组表达蛋白MEA具有良好的免疫反应性,建立的MEA-ELISA方法可用于HFRS患者血清特异性抗体的检测。

Objectives To express and purify a multi-epitope antigen（MEA）derived from the G2glycoprotein of Hantavirus and establish a method of MEA-ELISA to detect specific antibodies in serum from patients with hemorrhagic fever with renal syndrome（HFRS）.Methods MEA was expressed with a pET prokaryotic system and purified with nickel affinity chromatography.The immunoreaction specificity of MEA was investigated using Western blotting.A method of MEA-ELISA was established to detect specific antibodies in patients with HFRS.Results An MEA was successfully expressed with a pET prokaryotic expression system.The MEA protein had a molecular weight of 36.9ku,and the yield after purification was 0.15mg/ml.Western blot analysis indicated the target band of MEA was recognized by serum from patients with HFRS.MEA-ELISA results were positive for 120patients with HFRS at a rate of 90.83%and were negative for 60healthy individuals.Conclusion The recombinant protein MEA had good immunoreactive specificity.MEAELISA was able to detect specific antibodies in serum from patients with HFRS.