Affinity maturation of antibodies by combinatorial codon mutagenesis versus error-prone PCR.

ABSTRACT:

In vitro

affinity maturation of therapeutic monoclonal antibodies is commonly applied to achieve desired properties, such as improved binding kinetics and affinity. Currently there are no universally accepted protocols for generation of variegated antibody libraries or selection thereof. Here, we performed affinity maturation using a yeast-based single-chain variable fragment (scFv) expression system to compare two mutagenesis methods: random mutagenesis across the entire V(D)J region by error-prone PCR, and a novel combinatorial mutagenesis process limited to the complementarity-determining regions (CDRs). We applied both methods of mutagenesis to four human antibodies against well-known immuno-oncology target proteins. Detailed sequence analysis showed an even mutational distribu

SUBMITTER: Simons JF 

PROVIDER: S-EPMC7531523 | biostudies-literature | 2020 Jan-Dec

REPOSITORIES: biostudies-literature