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

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ROSTAMI B.; IRANI SH.; BOLHASSANI A.; Ahangari Cohan R.

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Journal: Pathobiology Research Year:2019 | Volume:22 | Issue:3 Page(s): 135-140

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Information Journal Paper

Title

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

Author(s)

ROSTAMI B. | IRANI SH. | BOLHASSANI A. | Ahangari Cohan R. | Issue Writer Certificate

Keywords

HIVQ3

NefQ1

Hsp20Q1

Prokaryotic SystemQ1

ImmunogenicityQ1

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 an immunomodulator or antigen carrier for subunit vaccine candidates. In the current study, the generation of Hsp20-Nef fusion protein was performed in E. coli expression system, and its Immunogenicity was evaluated in BALB/c mice. Materials and Methods At first, expression of Hsp20-Nef recombinant protein was studied in E. coli 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 kDa in 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 IgG as compared to other groups. Moreover, the Immunogenicity 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. coli Prokaryotic System and significantly induced antibody response against HIV-1 Nef antigen.

Cites

The Effects of Heat Shock Proteins on Delivery of HIV-1 Nef Antigen in Mammalian Cells

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APA: Copy

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. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), 22(3 ), 135-140. SID. https://sid.ir/paper/373590/en

Vancouver: Copy

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. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES)[Internet]. 2019;22(3 ):135-140. Available from: https://sid.ir/paper/373590/en

IEEE: Copy

B. ROSTAMI, SH. IRANI, A. BOLHASSANI, and R. Ahangari Cohan, “Expression of the Recombinant Hsp20-Nef Protein for Evaluation of its Immunogenicity against HIV-1 Nef using Indirect ELISA,” PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), vol. 22, no. 3 , pp. 135–140, 2019, [Online]. Available: https://sid.ir/paper/373590/en

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