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

Author(s):

Shekoufa Jahedian, Seyed Mehdi Sadat*, Gholam Reza Javadi and Azam Bolhassani   Pages 1 - 9 ( 9 )

Abstract:


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

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 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 anti-Nef antibody through western blotting. Data analyses showed that the protein has no considerable toxicity effect and has improved the IMT-P8 penetration rate in comparison with 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 into mammalian cell line and low toxicity for potential application as a vaccine against HIV-1.

Keywords:

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

Affiliation:

Department of Biology, Sciences and Research Branch, Islamic Azad University, Tehran, Department of Hepatitis, AIDS and Blood-borne diseases, Pasteur Institute of Iran. Tehran, Department of Biology, Sciences and Research Branch, Islamic Azad University, Tehran, Department of Hepatitis, AIDS and Blood-borne diseases, Pasteur Institute of Iran. Tehran



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