抗汉坦病毒NP scFv基因表达及生物活性分析

Gene Expression and Bio-Activity Analysis of scFv of Hantavirus NP

诱导已构建的重组质粒pGEX-6P-1-scFv原核表达抗汉坦病毒核衣壳蛋白单链抗体,并用酶免疫实验检测单链抗体生物活性。用IPTG诱导重组原核表达质粒pGEX-6P-1-scFv表达抗汉坦病毒NP单链抗体融合蛋白,经亲和层析纯化,并应用SDS-PAGE电泳检测单链抗体融合蛋白,应用酶免疫实验检测抗NP单链抗体生物学活性。SDS-PAGE电泳检测显示,原核重组质粒pGEX-6P-1-scFv已表达分子量约为56ku的单链抗体融合蛋白;酶免疫实验检测显示,单链抗体具有与汉坦病毒NP抗原特异性结合的生物学活性。结果表明,已构建的原核表达重组质粒pGEX-6P-1-scFv,能够成功表达具有与汉坦病毒NP抗原特异性结合生物学活性的单链抗体。

A constructed recombinant plasmid pGEX-6P-1- seFv was induced to express prokaryotie single-chain fragment of variable region （scFv） against nueleocapsid protein （NP） of Hantavirus and to test the bio-aetivity of scFv with immunoassay. IPTG was used to induce the recombinant that expressed prokaryotie plasmid pGEX-6P-1- seFv to express the fusion protein of NP seFv against Hantavirus. After purified by affinity and chromatography, the fusion seFv was tested with SDS-PAGE, bio-aetivity of anti-NP scFv was tested with immunoassay. The results of SDS-PAGE showed that prokaryotie recombinant plasmid pGEX-6P-1- seFv had expressed molecular mass of 56 ku of fusion protein of seFv, and immunoassay test showed that scFv possessed bio-aetivity of ability of specific binding to NP of Han- tavirus. The results showed that the prokaryotic recombinant plasmid pGEX-6P-1- seFv could successfully express the seFv possessing the ability of specific binding to NP of Hantavirus.