A stepwise mutagenesis approach using histidine and acidic amino acid to engineer highly pH-dependent protein switches.

ABSTRACT: Antibody-based drugs can be highly toxic, because they target normal tissue as well as tumor tissue. The pH value of the extracellular microenvironments around tumor tissues is lower than that of normal tissues. Therefore, antibodies that engage in pH-dependent binding at slightly acidic pH are crucial for improving the safety of antibody-based drugs. Thus, we implemented a stepwise mutagenesis approach to engineering pH-dependent antibodies capable of selective binding in the acidic microenvironment in this study. The first step involved single-residue histidine scanning mutagenesis of the antibody's complementarity-determining regions to prescreen for pH-dependent mutants and identify ionizable sensitive hot-spot residues that could be substituted by acidic amino acids to obtain pH-depen

SUBMITTER: Zou W 

PROVIDER: S-EPMC8686790 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature