Abstract
The small (S) envelope protein of the Hepatitis B Virus (HBV), HBV-S, has the unique ability to self-assemble into highly immunogenic subviral particles (SVPs), in the absence of other viral factors, in eukaryotic cells, including those of nonhepatic origin. This feature is currently exploited for generation of SVPs exposing heterologous epitopes on their surface that can be used as vaccine candidates to target various diseases. Here, we describe a simple and robust method for production of such chimeric HBV-S protein-based SVPs in transiently transfected HEK293T cells and purification from cell supernatants by ultracentrifugation on sucrose cushion and sucrose step gradients. The SVPs obtained by this methodology have been successfully used in immunogenicity studies in animal models.
Highlights
A variety of expression systems are available for production of protein antigens and vaccine development
The dimers spontaneously associate into 20 nm-diameter subviral particles (SVPs) that do not incorporate the viral capsid and genetic material and are secreted from cells, independent of virions (Fig.1) [5]
These virus-like particles (VLPs) are highly immunogenic, non-infectious and can be produced in large amounts in heterologous expression systems in the absence of any other viral components, which has led to their development into efficient and safe vaccines against Hepatitis B Virus (HBV) [6]
Summary
A variety of expression systems are available for production of protein antigens and vaccine development. The dimers spontaneously associate into 20 nm-diameter subviral particles (SVPs) that do not incorporate the viral capsid and genetic material and are secreted from cells, independent of virions (Fig.1) [5] These virus-like particles (VLPs) are highly immunogenic, non-infectious and can be produced in large amounts in heterologous expression systems in the absence of any other viral components, which has led to their development into efficient and safe vaccines against HBV [6]. These remarkable properties of the HBV-S protein have been exploited to generate chimeric SVPs carrying foreign and HBV-derived epitopes either fused or co-expressed with HBV-S [7–11]. This method is simple and scalable and may be applied to similar chimeric HBV particles displaying relevant immunogenic peptides derived from other pathogens of medical interest
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