Production and purification of chimeric HBc virus-like particles carrying influenza virus LAH domain as vaccine candidates

Andris Kazāks ,David J Rowlands,Alejandro Krimer,Benjamin A F Bláha ,Vincenzo Crescente,Olotu W Ogonah ,Sophie Farinelle,Alex Ramirez,Mapi Perez De Obanos,I-Na Lu,Tatjana Kazaka,Claude P Muller,Tarit Mukhopadhyay,Ināra Akopjana ,Anna Kirsteina,Nicola J Stonehouse,Velta Ose,Jānis Bogans ,Kaspars Tārs ,William Rosenberg

doi:10.1186/s12896-017-0396-8

Abstract

BackgroundThe lack of a universal influenza vaccine is a global health problem. Interest is now focused on structurally conserved protein domains capable of eliciting protection against a broad range of influenza virus strains. The long alpha helix (LAH) is an attractive vaccine component since it is one of the most conserved influenza hemagglutinin (HA) stalk regions. For an improved immune response, the LAH domain from H3N2 strain has been incorporated into virus-like particles (VLPs) derived from hepatitis B virus core protein (HBc) using recently developed tandem core technology.ResultsFermentation conditions for recombinant HBc-LAH were established in yeast Pichia pastoris and a rapid and efficient purification method for chimeric VLPs was developed to match the requirements for industrial scale-up. Purified VLPs induced strong antibody responses against both group 1 and group 2 HA proteins in mice.ConclusionOur results indicate that the tandem core technology is a useful tool for incorporation of highly hydrophobic LAH domain into HBc VLPs. Chimeric VLPs can be successfully produced in bioreactor using yeast expression system. Immunologic data indicate that HBc VLPs carrying the LAH antigen represent a promising universal influenza vaccine component.

Highlights

The lack of a universal influenza vaccine is a global health problem
Modification of the hepatitis B virus core protein (HBc) gene The insertion of heterologous sequences into viral structural genes generates chimeric virus-like particles (VLPs) that can be used as stable nanocontainers for diagnostics, vaccination and gene transfer purposes [11, 12]
We have demonstrated that the tandem core technology is a useful tool for incorporation of “difficult” sequences like the hydrophobic long alpha helix (LAH) domain into HBc VLPs while cloning into the classical, unmodified monomeric HBc gene results in insoluble products

Summary

Introduction

The lack of a universal influenza vaccine is a global health problem. Interest is focused on structurally conserved protein domains capable of eliciting protection against a broad range of influenza virus strains. A number of attempts have been made to Kazaks et al BMC Biotechnology (2017) 17:79 construct broadly protective candidate vaccines based on the conserved extracellular domain of matrix protein 2 (M2e) of influenza A viruses [6,7,8,9]. These M2e-based VLP vaccine candidates have been successfully tested in animal models, further efficacy studies are needed to demonstrate their protective potential in humans [1]. Many scientists believe that a universal and broad cross-protective influenza vaccine may require additional components One such potential candidate is the structurally conserved 55 amino acid-long alpha helix (LAH) of the HA stalk domain

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Conclusion