Implementing The Methodology To Generate Diagnostic And Therapeutic Antibodies

This research aimed to establish the methodology for developing monoclonal antibodies at The University of Texas at El Paso. Monoclonal antibodies are indispensable across multiple fields, including diagnostics, therapeutics, and biotechnology, yet there is a shortage of commercially available antibodies targeting specific proteins or possessing specialized characteristics. To address this gap, three models were selected: a peptide derived from the intracellular protein Schlafen 8 and two cell surface protein receptors: mouse vascular endothelial growth factor receptor 2 (VEGFR2) and human melanocortin receptor 4 (MC4R). These proteins present unique challenges in antibody generation, including issues of accessibility, expression, and specificity. Throughout this project, I developed expertise in various essential techniques, including immunization protocols, hybridoma generation, enzyme-linked immunosorbent assay (ELISA) development, western blotting, lentivirus production, generation of stable cell lines, flow cytometry, immunoprecipitation, gene cloning, and tissue culture techniques. This research lays the foundation for future antibody development efforts targeting proteins with complex characteristics.