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All Journal The Journal of Experimental Life Sciences (JELS)
Rohmah, Ilmiana Nurur
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Biochemistry, Genetics & Molecular Biology Immunology & microbiology Medicine & Pharmacology Neuroscience

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Evaluating SARS-CoV-2 Spike Protein Transfection in HEK-293T Cells for VLP Applications Rohmah, Ilmiana Nurur; Hafil Kusuma, Kavana; Ika Christina, Yuyun; Rizqi Dwijayanti, Dinia; Mustikaningtyas, Dewi; Widodo, Nashi; Sasmito Djati, Muhammad
The Journal of Experimental Life Science Vol. 14 No. 3 (2024)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2024.014.03.02

Abstract

The spike protein in SARS-CoV-2 works mainly to initiate infections, especially for binding to host cells and fusing with cell membranes. Characterizing SARS-CoV-2 spikes protein transfection cells could optimize the production of VLP in HEK-293T cells. Therefore, the present study aimed to investigate the characteristics of SARS-CoV-2 spike protein expression in transfected HEK-293T cells. Characterization of spike protein in transfected-HEK 293T cells was used to determine VLP formation in SARS-CoV-2. The study was conducted by isolating DNA and protein from transfected HEK 293T cells and analyzing spike expression using Polymerase Chain Reaction (PCR) and Flow Cytometry. The success of the SARS-CoV-2 spike protein transfection in PCR analysis has yet to be confirmed because DNA band smears do not specifically indicate the spike protein's size. However, the SARS-CoV-2 spike protein was successfully confirmed using flow cytometry with the percentage of spike-EGFP by 30%. Therefore, the present study confirmed the presence of spike protein in transfected HEK 293T cells by flow cytometry analysis. Keywords: Coronavirus, Characterization, DNA, Spike, Virus-Like Particle (VLP)
Corrigendum: Evaluating SARS-CoV-2 Spike Protein Transfection in HEK-293T Cells for VLP Applications Rohmah, Ilmiana Nurur; Marlita, Marlita; Kusuma, Kavana Hafil; Christina, Yuyun Ika; Dwijayanti, Dinia Rizqi; Mustikaningtyas, Dewi; Widodo, Nashi; Djati, M. Sasmito
The Journal of Experimental Life Science Vol. 15 No. 2 (2025)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2025.015.02.01

Abstract

The spike protein of SARS-CoV-2 is crucial for initiating infections by binding to host cells and mediating membrane fusion. In this study, HEK-293T cells were transfected with plasmids encoding three structural proteins of SARS-CoV-2, i.e., Spike (S), Membrane (M), and Envelope (E). This transfection enabled the formation of SARS-CoV-2 Virus-Like Particles (VLPs), which allows for safer studies of the virus and its proteins. To serve as a marker for expression, an enhanced green fluorescent protein (EGFP) was fused to the spike protein, resulting in a Spike-EGFP (S-EGFP) fusion protein. The characteristics of SARS-CoV-2 spike protein expression in the transfected HEK-293T cells were then investigated using polymerase chain reaction (PCR) and flow cytometry. The PCR analysis revealed non-specific DNA band smearing, which did not provide conclusive confirmation of spike protein expression. However, flow cytometry analysis demonstrated that approximately 30% of the transfected cells exhibited green fluorescence, indicating the expression of the Spike-EGFP fusion protein. These findings, obtained through flow cytometry, confirmed the successful spike protein expression in transfected HEK-293T cells.  
Co-Authors Dewi Mustikaningtyas Dinia Rizqi Dwijayanti Hafil Kusuma, Kavana Ika Christina, Yuyun Kusuma, Kavana Hafil Marlita Marlita, Marlita Moch Sasmito Djati Nashi Widodo Rizqi Dwijayanti, Dinia Yuyun Ika Christina