Adan Medina

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Faculty Mentor: Karl Dane Wittrup

Direct Supervisor: James A Van Deventer

Home University: Washington State University

Major: Chemical Engineering

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Biography

I am a senior at Washington State University, where I enjoy reading, flying paper/Styrofoam airplanes, and watching movies with my friends when I have spare time. My course of study is Chemical Engineering with an emphasis in microbiology. Although a bit unorthodox, these two fields intersect beautifully with my research interests in alternative energies (Biofuels). I chose Chemical engineering because I knew I could make the largest societal impact by decreasing humanity’s dependence on fossil fuels through research in alternative energies.

Abstract

Conversion of antibody fragments into antibody-like reagents

Many applications in basic science, diagnostics, and therapeutics rely on the use of antibodies or antibody-like reagents. Antibody fragments are central to many applications in the health sciences and are very easy to work with, but are not effective reagents for research without the corresponding constant region (Fc) of the antibody. Additionally, the process of isolating high-quality antibodies is very challenging and is often limited to experts. Thus, having access to a system capable of isolating antibody-like reagents with minimal difficulty is ideal. In this work, we focus on converting scFvs to scFv-Fcs in a yeast-based system suitable for the cell surface display and secretion of antibody-like reagents of interest. Antibody fragments were converted into scFv-Fcs using recombination in yeast to insert the DNA encoding an scFv into an Fc-encoding vector to create the gene coding for the scFv-Fc fusion protein. As test cases, we used previously isolated scFvs that recognize lysozyme and the neonatal Fc receptor (FcRn). We have used yeast surface display, immunoblots, and magnetic bead experiments to test for display and secretion of full-length, functional scFv-Fcs. Yeast surface display experiments indicate that comparable levels of scFv and scFv-Fc can be displayed on the yeast surface, while binding experiments indicate that scFvs are better binders then their scFv-Fc counterparts. Investigation of secreted samples of scFv-Fcs is being performed using immunoblots and magnetic bead experiments to test for full-length scFv-Fc secretion and for antigen binding function.