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

Curr HIV Res. 2020;18(5):315-323. doi: 10.2174/1570162X18666200612151925.

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

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS Vaccines / immunology*
  • Animals
  • Cell Line
  • Cell-Penetrating Peptides / genetics
  • Cell-Penetrating Peptides / metabolism*
  • HIV Infections / prevention & control*
  • HIV Infections / virology
  • HIV-1 / genetics
  • HIV-1 / immunology*
  • Humans
  • Mice
  • Recombinant Fusion Proteins
  • nef Gene Products, Human Immunodeficiency Virus / genetics
  • nef Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

  • AIDS Vaccines
  • Cell-Penetrating Peptides
  • Recombinant Fusion Proteins
  • nef Gene Products, Human Immunodeficiency Virus
  • nef protein, Human immunodeficiency virus 1