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
Show full item record
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
329
views
800
downloads
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

Suggestions

Development and immunological evaluation of SARS-CoV-2 virus-like particles (VLP) expressing the prefusion stabilized variants of the Spike protein
İpekoğlu, Emre Mert; Gürsel, Mayda; Department of Molecular Biology and Genetics (2022-1-24)
Emergence of the COVID-19 pandemic necessitated rapid development of highly effective vaccines against SARS-CoV-2. One adaptable strategy to produce a vaccine candidate against SARS-CoV-2 is based on the exploitation of the self- assembly feature of the structural proteins of SARS-CoV-2: Spike, Membrane, Envelope and Nucleocapsid, forming Virus-like particles (VLP). Our early studies focused on laboratory-scale production of SARS-CoV-2 VLPs in a mammalian host cell system based on transient expression of vi...
Two formulations of coronavirus disease-19 recombinant subunit vaccine candidate made up of S1 fragment protein P1, S2 fragment protein P2, and nucleocapsid protein elicit strong immunogenicity in mice
Özcengiz, Erkan; Keser, Duygu; Özcengiz, Gülay; Çelik, Gözde; ÖZKUL, AYKUT; İnçeh, Fatma Nur (2022-12-01)
Coronavirus disease (COVID-19) is ongoing as a global epidemic and there is still a need to develop much safer and more effective new vaccines that can also be easily adapted to important variants of the pathogen. In the present study in this direction, we developed a new COVID-19 vaccine, composed of two critical antigenic fragments of the S1 and S2 region of severe acute respiratory syndrome coronavirus 2 as well as the whole nucleocapsid protein (N), which was formulated with either alum or alum plus mon...
Effect of UV treatment for the removal of sulfonamide resistance gene during wastewater treatment
Bulut, Taliye; İçgen, Bülent (2018-04-27)
The dissemination of the antibiotic resistance genes (ARGs) is considered as one of the most important threats to public health. This dissemination also becomes a threat for water quality in surface waters and groundwater. Waters contaminated with ARGs can easily be incorporated into human life through several ways like drinking water, agriculture and animal husbandry. Sulfonamide resistance gene, one of the ARGs, in discharged waters increases gradually because the use of these drugs is highly common. Wast...
FACTORS THAT ARE ASSOCIATED WITH WILLINGNESS TO BE VACCINATED AND THE PERCEIVED EFFECTIVENESS OF VACCINES FROM ABROAD AND LOCAL VACCINES
Bilgen, Emine; Öner-Özkan, Bengi; Department of Psychology (2023-1)
The aim of this study is to examine the factors that predict people's vaccination decisions and the perceived effectiveness of vaccines developed and produced abroad and vaccines developed and produced in Turkey. 67.6% of respondents were willing to be vaccinated. The results showed that people who accepted vaccinations had higher levels of COVID-19-related anxieties, they reported more trust in vaccine companies and doctors, and scored low on trust in governmental institutions, but on the other hand, were ...
Identification of biomarkers associated with partial epithelial to mesenchymal transition in the secretome of slug over-expressing hepatocellular carcinoma cells
Karaosmanoglu, Oguzhan; Banerjee, Sreeparna; SİVAS, HÜLYA (Springer Science and Business Media LLC, 2018-08-01)
Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths worldwide. Complete epithelial to mesenchymal transition (EMT) has long been considered as a crucial step for metastasis initiation. It has, however, become apparent that many carcinoma cells can metastasize without complete loss of epithelial traits or with incomplete gain of mesenchymal traits, i.e., partial EMT. Here, we aimed to determine the similarities and differences between complete and partial EMT through over-expr...
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.
METU IIS - Integrated Information Service open@metu.edu.tr