Parallel Evolution of Antibody Affinity and Thermal Stability for Optimal Biotherapeutic Development

: Naive antibody libraries provide a rich resource for the identification of binding domains against targets of therapeutic interest. Being naive in nature means a lack in antigen bias, resulting in a breadth of diversity with respect to epitopes that can be successfully targeted. In combination with display-based technology platforms, selection strategies allow for the generation of ortholog cross-reactive binding domains which enable critical preclinical proof-of-concept studies. However, naive binding domains often suffer from low target affinity. In addition, construction of large naive libraries results in non-native pairing of heavy and light v-domains which can present a challenge to molecular stability. Here we describe effective methods for the parallel evolution of antibody affinity and thermal stability which couple mutant antibody library phage display with carefully designed selection strategies.