Title:
The Effect of Heterofunctional Microparticles on Identifying High Producing CHO cells
The Effect of Heterofunctional Microparticles on Identifying High Producing CHO cells
Author(s)
Aurelio, Caroline
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Abstract
Antibodies are produced by the human body in response to a foreign antigen such as a virus, parasite, and bacteria. Sometimes, the immune system fails to make enough antibodies to combat disease so man made monoclonal antibodies can be added to amplify immune response. The monoclonal antibodies are grown in host cells called CHO cells, or chinese hamster ovary cells, that are compatible with the growth of the target antigen and antibody pair. Monoclonal antibody treatments have been fabricated for diseases including Blood vessel tumor growth, Crohn's disease, Rheumatoid arthritis, sepsis and COVID-19. While monoclonal antibody drugs allow for very specific treatment and targeting of the disease, the process of growing the cell population and evaluating antibody production is very slow and burdensome. Traditional cell analysis methods including ELISA, or enzyme-linked immunosorbent assay, flow cytometry that measures antibody production through fluorescently stained antibodies, are both time consuming and cannot analyze cell secretion at the individual level. Therefore, determining a means by which the highest producing CHO cells can be identified and separated is imperative to streamline the drug production process. We created a heterofunctional microparticle called a Janus particle, that has targeting antibody on one side to bind the cell and protein G collection agent on the other to collect the released antibody. We cultured the CHO cells and incubated them with the fluorescent antigen and antibody combination, then added microparticles to the cell population at a microparticle:cell density of 100:1, ran the cells through flow cytometry and then separated the high and low secreting cells and ran traditional ELISA for verification. Through the success seen in this experiment, streamlining the production of monoclonal antibody drugs seems possible, whether it be through CHO or hybridoma cell hosts. Monoclonal antibodies have long term treatment applications including combating cancer, autoimmune diseases and infectious disease. In the future, a reagent with these microparticles can allow for separation of high producing cells that will allow for quicker drug clinical testing and fabrication.
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Date Issued
2021-12
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Text
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Undergraduate Thesis