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Updated: Aug 15, 2026
Isolation of Fidelity Variants of RNA Viruses and Characterization of Virus Mutation Frequency
Published on: June 16, 2011
Quantitative analysis of the effect of the mutation frequency on the affinity maturation of single chain Fv
P S Daugherty1, G Chen, B L Iverson
1Department of Chemical Engineering, Institute for Cell and Molecular Biology, University of Texas, Austin, TX 78712, USA.
Abstract:
Random mutagenesis and selection using phage or cell surface display provides an efficient method for affinity maturation of single chain Fv (scFv) antibodies, thereby improving function in various applications. To investigate the effects of mutation frequency on affinity maturation, error-prone PCR was used to generate libraries containing an average (m) of between 1.7 and 22.5 base substitutions per gene in a high affinity scFv antibody that binds to the cardiac glycoside digoxigenin. The scFv antibody libraries were displayed on Escherichia coli, and mutant populations were analyzed by flow cytometry. At low to moderate mutation frequencies with an average mutation rate of m
