Tarbiat Modares University Pathobiology Research 2538-3000 22 3 2019 07 01 Expression of the Recombinant Hsp20-Nef Protein for Evaluation of its Immunogenicity against HIV-1 Nef using Indirect ELISA بیان پروتئین نوترکیب Hsp20-Nef به‌منظور ارزیابی ایمنوژنیسیتی آن در مقابل HIV-1 Nef توسط الایزای غیرمستقیم 135 140 19353 EN B. Rostami Biology Department, Sciences & Research Branch, Islamic Azad University, Tehran, Iran Sh. Irani Biology Department, Sciences & Research Branch, Islamic Azad University, Tehran, Iran A. Bolhassani Hepatitis & AIDS Department, Pasteur Institute of Iran, Tehran, Iran R. Ahangari Cohan Pilot Nano-Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran Journal Article Aims: Nef protein has been considered as an attractive target for the development of therapeutic HIV-1 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 Hsp20-Nef fusion protein was performed in E. , and in BALB/c mice. Materials and Methods: At first, of Hsp20-Nef recombinant protein E. BL21 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-1 Nef antigen using indirect ELISA in mice. Findings: Our data showed a clear band of ~1230bp related to Hsp20-Nef fusion on agarose gel indicating the correct gene cloning in pET28a vector. The expression of Hsp20-Nef protein was confirmed as a clear band of ~47 SDS-PAGE and western blotting. In the immunological assay, the Hsp20-Nef protein and also the Nef protein emulsified with Freund’s adjuvant significantly enhanced the level of total compared to other groups. Moreover, of Hsp20-Nef was higher than Freund’s adjuvant/Nef in protein regimens (p<0.05). Conclusion: The Hsp20-Nef fusion protein was effectively expressed in E. and significantly induced antibody response against HIV-1 Nef antigen. https://mjms.modares.ac.ir/article_19353_35afcb39dad0fd02235dedfab3beb4ab.pdf