Abstract
Virus-like particles (VLPs) constitute a promising approach to recombinant vaccine development. They are robust, safe, versatile and highly immunogenic supra-molecular structures that closely mimic the native conformation of viruses without carrying their genetic material. HIV-1 Gag VLPs share similar characteristics with wild-type severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus, making them a suitable platform for the expression of its spike membrane protein to generate a potential vaccine candidate for COVID-19. This work proposes a methodology for the generation of SARS-CoV-2 VLPs by their co-expression with HIV-1 Gag protein. We achieved VLP functionalization with coronavirus spike protein, optimized its expression using a design of experiments (DoE). We also performed the bioprocess at a bioreactor scale followed by a scalable downstream purification process consisting of two clarifications, an ion exchange and size-exclusion chromatography. The whole production process is conceived to enhance its transferability at current good manufacturing practice (cGMP) industrial scale manufacturing. Moreover, the approach proposed could be expanded to produce additional Gag-based VLPs against different diseases or COVID-19 variants.
Highlights
COVID-19 is a disease caused by severe acute respiratory syndrome coronavirus2 (SARS-CoV-2)
To produce SARS-CoV-2 spike Gag::eGFP Virus-like particles (VLPs) (S-VLPs), cells were co-transfected with plasmids pGag::eGFP and pSpike, using PEI as the transfection reagent
In order to track the HIV-1 Gag polyprotein expression and characterize S- and G-VLPs, Gag was fused in frame with eGFP, as previously reported [26]
Summary
It emerged in December 2019 in Wuhan and since has spread around the globe causing a pandemic that had devastating health and economic consequences worldwide [1,2]. An enormous effort made by the scientific community resulted in more than 300 new vaccine candidates in less than a year since the outbreak, some of them being approved for emergency use [3], as well as the development of diagnosis methods for its detection [4] and treatment [5]. There are still some questions left to answer, like how long the immune memory lasts, the protective effect that current approved vaccines generate against emerging SARS-CoV-2 variants, or if it is possible to generate fully prophylactic vaccines against this new coronavirus [8]
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