Honors Theses

Date of Award

Spring 5-9-2024

Document Type

Undergraduate Thesis

Department

Biology

First Advisor

Wayne Gray

Second Advisor

Colin Jackson

Third Advisor

Eden Tanner

Relational Format

Dissertation/Thesis

Abstract

In 2020, the world was introduced to COVID-19, a strain of Coronavirus that quickly caused a pandemic at the global scale. COVID-19 was reported to cause 5.94 million deaths between 2020-2021, and it continues to prevail amongst society in 2024. Many methods of preventing COVID-19 infection arose throughout this time period; however, mRNA vaccines against the SARS-CoV-2 spike (S) protein became the most effective way to protect the population from COVID-19’s destructive reign.

Although society has made significant scientific strides in Coronavirus testing and vaccination efforts, there are many improvements that must be made to better protect our society from COVID-19. This thesis intends to investigate the immunoblot assay as an antibody test and how it can be utilized to more accurately detect COVID-19 infection in contrast to the ELISA assay, which provides less consistent rates of accuracy. The protein of interest, the SARS-CoV-2 nucleocapsid (N) protein expressed in E.Coli, was isolated using transformation and column purification methods. Protein electrophoresis and chemiluminescent western blot analysis were used to ensure the protein’s purification. COVID-19 vaccinated patient serum and infected patient serum were analyzed using these methods to test for the presence of antibodies to the N protein. The results from these immunoblot assays were contrasted with results from similar assays utilizing the SARS-CoV-2 S protein. The SARS-CoV-2 N protein has proven to be more stable and less resistant to mutations across all new variants of COVID-19, as opposed to the virus’s S protein. By highlighting the presence of antibodies to the nucleocapsid in patient serum using immunoblot antibody testing, we hope to emphasize the future possibility of creating a one-time mRNA vaccine against the SARS-CoV-2 nucleocapsid protein.

Creative Commons License

Creative Commons Attribution 4.0 International License
This work is licensed under a Creative Commons Attribution 4.0 International License.

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