Evalynn Ellison, ‘25 chemical engineering, secured the title of top junior presenter in the engineering, physics, and mathematics category for presenting the poster based on her summer research project titled, "In-Frame Cloning of Chimeric Antigen Receptors to Improve Cancer Therapeutic Discovery" at the Annual Biomedical Research Conference for Minority Students (ABRCMS) conference held in November 2023.
Poster title: In-Frame Cloning of Chimeric Antigen Receptors to Improve Cancer Therapeutic Discovery
Authors: Evalynn Ellison (1), Azady Pirhanov (2,3), and Brandon DeKosky (2,3)
1Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County (UMBC)
2Department of Chemical Engineering, Massachusetts Institute of Technology (MIT)
3The Ragon Institute of MGH, MIT, and Harvard
Abstract: Chimeric Antigen Receptors (CARs) are being used to create promising cancer immunotherapies; however, high-throughput CAR discovery remains a challenge due to the many possible proteins that must be screened for activity. In particular, creating very large and diverse CAR gene libraries for effective CAR discovery has been a limitation. This research project focused on developing and optimizing a system to identify productively inserted CAR genes after cloning into expression vectors. To accomplish this, we developed a high-throughput in-frame CAR clone identification strategy based on a fluorescent protein mCherry assay. The changes in fluorescent reporter mCherry expression levels would signal that a potential CAR was expressed in-frame. After cloning test genes, samples were analyzed by flow cytometry to measure the level of fluorescence and percentage of cells expressing fluorescent protein mCherry. We concluded that the designed mCherry-based fluorescent reporter system was sensitive and specific, making it a successful validator of cloned CAR genes. These CAR platforms can be used for cancer therapeutic discovery toward cell-based immunotherapies, such as CAR-Ts. Our new approach to identify CAR candidates will make therapeutic development more streamlined and efficient.
Photo credit: Evalynn Ellison