Abstract
BackgroundDevelopment 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 workThe 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 resultsGel.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).ConclusionGel.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.
Highlights
Egypt has the highest prevalence of Hepatitis C Virus (HCV) infection in the world, making it one of the major public health challenges facing the country
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
NS2 gene was conjugated to gelatin nanoparticles and a successful conjugation was confirmed by labeling and imaging using Confocal Laser Scanning Microscope (CLSM)
Summary
Egypt has the highest prevalence of Hepatitis C Virus (HCV) infection in the world, making it one of the major public health challenges facing the country. As prevention is better than treatment, and to avoid the expected mortality from HCV-related cirrhosis or HCC in the decade; an effective vaccine against HCV should be developed [11]. For this purpose; nonstructural (NS2) protease domain, the most genetically conserved viral antigen among HCV genotypes, has been used for the induction of cellular immunity in animal models in different vaccine studies [12], this might help in the design of a vaccine against the predominant genotype 4a. Editor: Abhijit De, Advanced Centre for Treatment Research and Education in Cancer, INDIA
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