Intellectual Merit: Antibodies play an important role in the immune response by recognizing foreign molecules with high affinity and specificity. It is these same properties that make them excellent affinity reagents in fields ranging from basic research to diagnostics and nanotechnology. While efforts to obtain high affinity antibodies against almost any molecule of interest are now relatively straight-forward, methods to control and regulate antibody binding have not been well explored. Regulation of protein interactions is quite common in biology and frequently involves additional equilibria that are linked to protein binding. The research objective of this CAREER project is to study single domain antibody fragments as a vehicle for understanding and applying linked equilibria in biomolecular systems. By combining recent advances in protein engineering techniques with biophysical and structural analysis, this research will explore several methods to engineer reversible control of antibody interactions through the introduction of linked equilibria. Several approaches will be explored to either reduce or enhance antibody binding under defined conditions. The resulting knowledge, methods, and reagents will help stimulate new directions in research and biotechnology by providing unique routes to control protein interactions. These results will also be of fundamental interest to researchers interested in protein design, structure/function relationships, and computational chemistry. Furthermore, these results will help improve the general understanding and teaching of molecular regulation in biology.
Broader Impacts: The educational component complements the research aims and serves to enhance training at Northern Illinois University (NIU) and the regional community. To enhance the undergraduate curriculum at NIU, an upper-level undergraduate biochemistry lab will be re-designed to incorporate inquiry-based learning approaches. These training tools will be further developed through the initiation of a web-based wiki project to serve as a training resource for the larger community of biochemical instructors and students. To help attract and retain student interest in the sciences at NIU, a new course will be developed that is aimed at exposing undergraduate students to cutting edge research at the interface of biology and chemistry. In collaboration with NIU STEM Outreach, this course will also be adapted to summer workshops for pre-college students and secondary school instructors to help increase scientific literacy. This CAREER project also includes partnerships with NIU's STEM Outreach and Latino Resource Center to provide opportunities for high school students and underrepresented undergraduate students, respectively, to perform research in the PI's laboratory.
