The V_H framework region 1 as a target of efficient mutagenesis for generating a variety of affinity-matured scFv mutants.

ABSTRACT: In vitro affinity-maturation potentially generates antibody fragments with enhanced antigen-binding affinities that allow for developing more sensitive diagnostic systems and more effective therapeutic agents. Site-directed mutagenesis targeting "hot regions," i.e., amino acid substitutions therein frequently increase the affinities, is desirable for straightforward discovery of valuable mutants. We here report two "designed" site-directed mutagenesis (A and B) targeted the N-terminal 1-10 positions of the V_H framework region 1 that successfully improved an anti-cortisol single-chain Fv fragment (K_a, 3.6 × 10⁸ M^-1). Mutagenesis A substituted the amino acids at the position 1-3, 5-7, 9 and 10 with a limited set of substitutions to generate only 1,

SUBMITTER: Kiguchi Y 

PROVIDER: S-EPMC8050046 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature