Rostami, B., Irani, Sh., Bolhassani, A., Ahangari Cohan, R. (2019). Expression of the Recombinant Hsp20-Nef Protein for Evaluation of its Immunogenicity against HIV-1 Nef using Indirect ELISA. , 22(3), 135-140.
B. Rostami; Sh. Irani; A. Bolhassani; R. Ahangari Cohan. "Expression of the Recombinant Hsp20-Nef Protein for Evaluation of its Immunogenicity against HIV-1 Nef using Indirect ELISA". , 22, 3, 2019, 135-140.
Rostami, B., Irani, Sh., Bolhassani, A., Ahangari Cohan, R. (2019). 'Expression of the Recombinant Hsp20-Nef Protein for Evaluation of its Immunogenicity against HIV-1 Nef using Indirect ELISA', , 22(3), pp. 135-140.
Rostami, B., Irani, Sh., Bolhassani, A., Ahangari Cohan, R. Expression of the Recombinant Hsp20-Nef Protein for Evaluation of its Immunogenicity against HIV-1 Nef using Indirect ELISA. , 2019; 22(3): 135-140.

Expression of the Recombinant Hsp20-Nef Protein for Evaluation of its Immunogenicity against HIV-1 Nef using Indirect ELISA

Pathobiology Reserach
Article 4, Volume 22, Issue 3, 2019, Pages 135-140    PDF (765.96 K)
Document Type: Original Research
Authors
B. Rostami1; Sh. Irani1; A. Bolhassani* 2; R. Ahangari Cohan3
1Biology Department, Sciences & Research Branch, Islamic Azad University, Tehran, Iran
2Hepatitis & AIDS Department, Pasteur Institute of Iran, Tehran, Iran
3Pilot Nano-Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran
Abstract
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.
Keywords
HIV; Nef; Hsp20; Bacterial expression system; Immunogenicity
References
Statistics
Article View: 104
PDF Download: 61
Statistics
Number of Journals45
Number of Issues2,171
Number of Articles24,674
Article View24,443,756
PDF Download17,553,707