A CURE for Cloning: Implementing a Modular Approach to Cloning Recombinant Antibodies in an Undergraduate Teaching Lab

Abstract

An undergraduate course‐based research experience (CURE) is a high impact practice which provides genuine research experiences to a large and diverse student population. We have implemented a six‐week research module involving the cloning and expression of therapeutic antibodies as the culmination of a senior level Biochemistry lab course. We have designed a cloning system that takes advantage of the modular domain structure of antibodies to facilitate changes to an antibody’s affinity, isotype, or allotype. Specifically, our system was developed for readily swapping the antigen‐binding N‐terminal sequences of different antibodies into a set of vectors encoding the conserved regions of the heavy and light chains, all in the background of a strong mammalian expression vector. Gene fragments corresponding to the constant regions of each immunoglobulin chain, as well as the antigen‐binding regions of unique antibodies were designed, and produced by gene synthesis. Various iterations of the module were tested over four different semesters with high levels of success in terms of student engagement and research goals. The resulting plasmid constructs are now being used by our research lab as part of the investigation of the nature of antibody recognition of cancer‐specific Mucin antigens. This project allowed our senior undergraduates to contribute to an active research program, while earning credit toward a required course. It exposed them to modern cloning methodologies, bioinformatics tools, cell culture techniques and participation in a local research conference. It is also a readily scalable project as only the variable regions of new antibodies of interest need be synthesized.