Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene

PLoS One. 2017 Jul 26;12(7):e0181723. doi: 10.1371/journal.pone.0181723. eCollection 2017.

Abstract

Background: Development of an effective non-viral vaccine against hepatitis C virus infection is of a great importance. Gelatin nanoparticles (Gel.NPs) have an attention and promising approach as a viable carrier for delivery of vaccine, gene, drug and other biomolecules in the body.

Aim of work: The present study aimed to develop stable Gel.NPs conjugated with nonstructural protein 2 (NS2) gene of Hepatitis C Virus genotype 4a (HCV4a) as a safe and an efficient vaccine delivery system.

Methods and results: Gel.NPs were synthesized and characterized (size: 150±2 nm and zeta potential +17.6 mv). NS2 gene was successfully cloned and expressed into E. coli M15 using pQE-30 vector. Antigenicity of the recombinant NS2 protein was confirmed by Western blotting to verify the efficiency of NS2 as a possible vaccine. Then NS2 gene was conjugated to gelatin nanoparticles and a successful conjugation was confirmed by labeling and imaging using Confocal Laser Scanning Microscope (CLSM). Interestingly, the transformation of the conjugated NS2/Gel.NPs complex into E. coli DH5-α was 50% more efficient than transformation with the gene alone. In addition, conjugated NS2/Gel.NPs with ratio 1:100 (w/w) showed higher transformation efficiency into E. coli DH5-α than the other ratios (1:50 and 2:50).

Conclusion: Gel.NPs effectively enhanced the gene delivery in bacterial cells without affecting the structure of NS2 gene and could be used as a safe, easy, rapid, cost-effective and non-viral vaccine delivery system for HCV.

MeSH terms

  • Drug Delivery Systems / methods
  • Gelatin / chemistry*
  • Genotype
  • Hepacivirus / genetics
  • Hepacivirus / metabolism*
  • Hepatitis C / immunology
  • Hepatitis C / prevention & control
  • Hepatitis C / virology
  • Humans
  • Microscopy, Confocal
  • Microscopy, Electron, Transmission
  • Nanoparticles / administration & dosage
  • Nanoparticles / chemistry*
  • Nanoparticles / ultrastructure
  • Particle Size
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Spectrophotometry, Ultraviolet
  • Viral Hepatitis Vaccines / administration & dosage
  • Viral Hepatitis Vaccines / chemistry
  • Viral Hepatitis Vaccines / immunology
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / immunology
  • Viral Nonstructural Proteins / metabolism*
  • X-Ray Diffraction

Substances

  • NS2 protein, Hepatitis C virus
  • Recombinant Proteins
  • Viral Hepatitis Vaccines
  • Viral Nonstructural Proteins
  • Gelatin

Grants and funding

The authors received no specific funding for this work.