HIV-1Gp41Recombinant Antigen: Expression In E.Coli And Immunoreactivity Analysis

Enzyme-Linked Immuno Sorbent Assay (ELISA) is still a mainstream clinical screening method for the detection of HIV (Human Immunodeficiency Virus) infection, due to its high testing throughput, low-cost and easy to operate. This study aims to express HIV-1transmembrane glycoprotein gp41, which has fine immunoreactivity, in prokaryotic system of E.coli.The study seleted DNA sequences coding major epitopes of HIV-1gp41as template, designed suitable primers pair, and then amplified target genes through PCR. The genes, after digested by BamH I and Xho I, inserted into plasmid vector E3. Then the joint product was transported into cloning host DH5a, from which the recombinant plasmid was extracted and then further transpoted into expressive host BL21(DE3). After being induced with IPTG, the recombinant protein was obtained. Due to the His-Tag the protein contained, it was purified with Nif-chelated affinity chromatopraphy. After that, the recombinant antigen was labeled with HRP, and antigenicity was analysed by detecting various specimens through double-antigen sandwich ELISA.In this study, HIV-1transmembrane glycoprotein gp41was successfully expressed in E.coli system. This recombinant antigen that contained major epitopes of HIV-1gp41was further used as a raw material, to develop HIV-ELISA kit, for the detection of HIV antibody in human blood. The antigenicity and the immunoreactivity of the recombinant protein of HIV-1gp41was analysed. The comprehansive evaluation of its sensitivity, specificity and stability was performed by comparing with several commercially available ELISA kits of good brand. The results showed a preferable performance of the kit that contained this recombinant antigen, this may partially lead to a commercial utilization of the antigen in potential HIV antibody detection kit.