Expression of the Recombinant Hsp۲۰-Nef Protein for Evaluation of its Immunogenicity against HIV-۱ Nef using Indirect ELISA

Aims: Nef protein has been considered as an attractive target for the development of therapeutic HIV-۱ vaccine. Furthermore, strong immunological properties of heat shock proteins (HSPs) led to their use as for subunit vaccine candidates. In the current study, the generation of Hsp۲۰-Nef fusion protein was performed in E. , and in BALB/c mice. Materials and Methods: At first, of Hsp۲۰-Nef recombinant protein E. BL۲۱ and Rosetta strains by SDS-PAGE and western blotting using anti-Nef monoclonal antibody. Then, the recombinant protein was purified by a reverse staining method. Finally, its potency was evaluated to elicit antibody response against HIV-۱ Nef antigen using indirect ELISA in mice. Findings: Our data showed a clear band of ~۱۲۳۰bp related to Hsp۲۰-Nef fusion on agarose gel indicating the correct gene cloning in pET۲۸a vector. The expression of Hsp۲۰-Nef protein was confirmed as a clear band of ~۴۷ SDS-PAGE and western blotting. In the immunological assay, the Hsp۲۰-Nef protein and also the Nef protein emulsified with Freund’s adjuvant significantly enhanced the level of total compared to other groups. Moreover, of Hsp۲۰-Nef was higher than Freund’s adjuvant/Nef in protein regimens (p<۰.۰۵). Conclusion: The Hsp۲۰-Nef fusion protein was effectively expressed in E. and significantly induced antibody response against HIV-۱ Nef antigen.

کلیدواژه ها:

HIV ، Nef ، Hsp۲۰ ، Bacterial expression system ، Immunogenicity ، ویروس HIV ، Nef ، Hsp۲۰ ، سیستم پروکاریوتی ، ایمنوژنیسیتی