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Unleasing Fas ligand: bacterial expression, purification, and functional analysis in melanoma and cervical cancer apoptosis : a thesis in Biomedical Engineering and Biotechnology
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Unleasing Fas ligand: bacterial expression, purification, and functional analysis in melanoma and cervical cancer apoptosis : a thesis in Biomedical Engineering and Biotechnology

Saadi Toni Saade
Master of Science (MS), University of Massachusetts Dartmouth
2024
DOI:
https://doi.org/10.62791/20528

Abstract

The use of Fas ligand could revolutionize the way that we target malignancies. Cancer has long been a fierce enemy; in the US, two million people received a cancer diagnosis in 2022, and 609,000 people lost their lives to the disease. Therefore, one of the main objectives in the field of biotechnology is the development of novel techniques to target and eliminate cancer cells. Fas ligand, an essential component of programmed cell death, has the ability to eradicate specific cancer cells, making it a prospective therapeutic candidate. The project entails cloning the Fas ligand gene molecularly into an expression vector for e. Coli and then stimulating the production of the protein in bacterial cultures. Affinity chromatography is applied to modified e. Coli strains using the GST-tag to obtain a highly concentrated and bioactivate Fas ligand. To assess the therapeutic potential, the purified product is applied to A375 melanoma cells and HeLa cell cultures with a determination of impact on cell viability and apoptosis. This would provide insight into the feasibility of using e. Coli production systems to produce effective therapeutics in targeting cancer.
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