Volume 22, Issue 3 (2019)
mjms 2019, 22(3): 135-140 |
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Rostami B, Irani S, Bolhassani A, Ahangari Cohan R. Expression of the Recombinant Hsp20-Nef Protein for Evaluation of its Immunogenicity against HIV-1 Nef using Indirect ELISA. mjms 2019; 22 (3) :135-140
URL: http://mjms.modares.ac.ir/article-30-25213-en.html
URL: http://mjms.modares.ac.ir/article-30-25213-en.html
1- Biology Department, Sciences & Research Branch, Islamic Azad University, Tehran, Iran
2- Hepatitis & AIDS Department, Pasteur Institute of Iran, Tehran, Iran ,azam_bolhassani@yahoo.com
3- Pilot Nano-Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran
2- Hepatitis & AIDS Department, Pasteur Institute of Iran, Tehran, Iran ,
3- Pilot Nano-Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran
Abstract: (7547 Views)
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.
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.
Article Type: Original Research |
Subject:
Molecular Biology
Received: 2018/09/17 | Accepted: 2019/02/7
Received: 2018/09/17 | Accepted: 2019/02/7
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