PRODUCTION OF VIRUS LIKE PARTICLES EXPRESSING ALPHA (B.1.1.7) OR DELTA PLUS (B.1.617.2.1) SPIKE VARIANTS AND DEVELOPMENT OF A PSEUDOTYPED VIRUS NEUTRALIZATION ASSAY

2022-2-10
Güvençli, Neşe
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Vaccines are one of the most important public health interventions to combat the spread of infectious diseases. All currently approved COVID-19 vaccines target Spike antigen derived from the authentic Wuhan strain. However, with the emergence of new variants effectiveness of the current vaccines declined, commensurate to their ability to generate neutralizing antibodies against some variants of concern (VOC). Since new VOCs are predicted to emerge the next 2-4 years, variant specific vaccines are urgently needed. In this study, we aimed to design vaccine antigens against Delta plus (B 1.617.2.1) and Alpha (B.1.1.7) variants of concern by exploiting the Virus Like Particle (VLP) vaccine platform. Consequently, these variant specific VLPs were formed by assembling the Membrane (M), Nucleocapsid (N), Envelope (E) proteins of the authentic virus and Spike (S) proteins bearing mutations of Delta plus (B 1.617.2.1) and Alpha (B.1.1.7) variants of concern. For this, the M and N genes were cloned into the pVitro1 expression vector, whereas variant specific Spike genes were cloned together with the Envelope gene into the pVitro2 expression vector. Transfection of these vectors to Hek293 suspension cells resulted in VLP accumulation in the supernatant. Herein, we demonstrate the feasibility of producing variant spike expressing VLPs derived from Alpha (B.1.1.7) and Delta plus (B.1.617.2.1) VOCs. Furthermore, we established a pseudotyped virus neutralization assay and tested its utility using sera taken from volunteers and mice immunized with Delta plusS-6p-VLP or AlphaS-6p-VLP vaccines.
SARS-CoV-2, Vaccine, Variants of Concern, Virus Like Particle, Neutralization Assay
https://hdl.handle.net/11511/96705
Graduate School of Natural and Applied Sciences, Thesis

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Citation Formats
N. Güvençli, “PRODUCTION OF VIRUS LIKE PARTICLES EXPRESSING ALPHA (B.1.1.7) OR DELTA PLUS (B.1.617.2.1) SPIKE VARIANTS AND DEVELOPMENT OF A PSEUDOTYPED VIRUS NEUTRALIZATION ASSAY,” M.S. - Master of Science, Middle East Technical University, 2022.
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