In Silico Design and Production of a Multiepitopic E2 Glycoprotein based Vaccine Against HCV Genotype 3a

Abstract

The infection caused by Hepatitis C virus (HCV) continues to pose a substantial global health concern, exerting a notable influence on the well-being of the general population. Once the HCV enters the body it starts affecting the liver causing acute hepatitis and if left untreated for a long time it can progressively lead to liver cancer. Despite the availability of direct acting antivirals there are high chances of reinfection which creates a crucial gap in preventative healthcare, particularly in areas where specific HCV genotypes, such as genotype 3a, are predominant. The goal set by WHO is to eradicate HCV infections by 2030. The major reason behind the unavailability of an HCV vaccine is its variable genome. E2 is the HCV receptor-binding protein and, consequently, the main target for bnAbs. However, a safe, prophylactic multivalent polypeptide vaccine with conserved epitopes is the most crucial need of the hour to globally eliminate HCV as development of broadly neutralizing antibodies against the envelop 2 glycoprotein of HCV is quite laborious owing to its conformational flexibility. In this study, using in silico approach conserved antigenic epitopes of E2 glycoprotein will be identified based on predictions, demonstrating B cell activation, T cell activation, and production of IFNγ, against HCV subtype 3a. The epitopes will be modeled into a vaccine construct using GPGPG linkers. To augment the immunogenicity of the construct, an adjuvant known as β-Defensin 2 will be included at the N-terminal region. Molecular docking analysis will indicate a strong contact between vaccine and its potential receptors. Immune simulations will indicate a robust immune response by minimizing the escape of HCV from neutralizing antibodies. The vaccine construct will be cloned into PET-28a(+) expression vector and subsequently be expressed in Escherichia coli BL21 bacteria. A rigorous purification technique will be devised in order to extract the recombinant multiepitopic protein, ensuring its purity and structural integrity. Hence, this multiepitopic candidate vaccine will show potential as a preventative and therapeutic tool, offering hope in reducing the burden of HCV infections on global public health.