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Current HIV Research

Author(s): Shekoufa Jahedian, Seyed Mehdi Sadat*, Gholam Reza Javadi and Azam Bolhassani

DOI: 10.2174/1570162X18666200612151925

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Production and Evaluation of the Properties of HIV-1-Nef-MPER-V3 Fusion Protein Harboring IMT-P8 Cell Penetrating Peptide

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Abstract

Background: Finding a safe and effective vaccine for HIV-1 infection is still a major concern.

Objective: This study aimed to design and produce a recombinant Nef-MPER V3 protein fused with IMT-P8 using E. coli expression system to provide a potential HIV vaccine with high cellular penetrance.

Methods: After synthesizing the DNA sequence of the fusion protein, the construct was inserted into the pET-28 expression vector. The recombinant protein expression was induced using 1 mM IPTG and the product was purified through affinity chromatography. Characterization of cellular delivery, toxicity and immunogenicity of the protein was carried out.

Results: The recombinant protein was expressed and confirmed by the anti-Nef antibody through western blotting. Data analyses showed that the protein possessed no considerable toxicity effect and has improved the IMT-P8 penetration rate in comparison to a control sample. Moreover, the antigen immunogenicity of the protein induced specific humoral response in mice.

Conclusion: It was concluded that IMT-P8-Nef-MPER-V3 fusion protein has a high penetrance rate in mammalian cell line and low toxicity, thus it can be potentially considered as a vaccine against HIV-1.

Keywords: HIV-1, Recombinant protein, Escherichia coli, Nef-MPER-V3, Cell penetrating peptide, IMT-P8.

Graphical Abstract

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