Expression and purification of recombinant HIV-1 gp41 protein and determination of its immunoreactivity

Objective Expressions of HIV-1 gp41 antigen for in-vitro immunoassay.Methods HIV-1 gp41 gene was amplified by PCR by using the plasmid X1 as a template,and then inserted into the plasmid E1 to construct recombinant plasmids E1-1.The expression of the recombinant plasmid in competent E.coli cell Bl21DE3 was induced with IPTG and analyzed with SDS-PAGE.After being purified by Ni-NTA affinity chromatography,the recombinant protein was further evaluated with ELISA for their functional characteristics.Results The result from SDS-PAGE showed expression of E1-1 in the competent E.coli cell.The purity of the purified recombinant protein was over 95%,and the protein has excellent immunoreactivity and specificity with ELISA.Conclusions Recombinant protein E1-1 can be expressed with a high immunoactivity,which can be qualified for HIV ELISA production.