Apr 28, 2020

In vitro Evolution of Antibody Affinity via Insertional Mutagenesis Scanning of an Entire Antibody Variable Region

BioRxiv : the Preprint Server for Biology
K. SkamakiFlorian Hollfelder


We report the first systematic combinatorial exploration of affinity enhancement of antibodies by insertions and deletions (InDels). Transposon-based introduction of InDels via the method TRIAD was used to generate large libraries with random in-frame InDels across the entire scFv gene that were further recombined and screened by ribosome display. Knowledge of potential insertion points from TRIAD libraries formed the basis of exploration of length and sequence diversity of novel insertions by insertional-scanning mutagenesis (ISM). An overall 256-fold affinity improvement of an anti-IL-13 antibody BAK1 as a result of InDel mutagenesis and combination with known point mutations validates this approach and suggests that the results of this InDel approach and conventional exploration of point mutations can synergize to generate antibodies with higher affinity.

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Mentioned in this Paper

Biochemical Pathway
MECP2 wt Allele
Methyl-CpG-Binding Protein 2
Regulation of Biological Process
Gene Expression
MECP2 gene
Study of Epigenetics
Binding Protein

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