Facile Affinity Maturation of Antibody Variable Domains Using Natural Diversity Mutagenesis.

ABSTRACT: The identification of mutations that enhance antibody affinity while maintaining high antibody specificity and stability is a time-consuming and laborious process. Here, we report an efficient methodology for systematically and rapidly enhancing the affinity of antibody variable domains while maximizing specificity and stability using novel synthetic antibody libraries. Our approach first uses computational and experimental alanine scanning mutagenesis to identify sites in the complementarity-determining regions (CDRs) that are permissive to mutagenesis while maintaining antigen binding. Next, we mutagenize the most permissive CDR positions using degenerate codons to encode wild-type residues and a small number of the most frequently occurring residues at each CDR position based on natural

SUBMITTER: Tiller KE 

PROVIDER: S-EPMC5591402 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature