In vitro Evolution of Antibody Affinity via Insertional Mutagenesis Scanning of an Entire Antibody Variable Region
作者： Kalliopi Skamaki , Stephane Emond , Matthieu Chodorge , John Andrews , D. Gareth Rees
DOI: 10.1101/2020.04.26.062786
关键词: Gene 、 Biology 、 Transposable element 、 Insertional mutagenesis 、 Computational biology 、 Ribosome display 、 Systematic evolution of ligands by exponential enrichment 、 Point mutation 、 Mutagenesis (molecular biology technique) 、 Indel
摘要: We report the first systematic combinatorial exploration of affinity enhancement antibodies by insertions and deletions (InDels). Transposon-based introduction InDels via method TRIAD was used to generate large libraries with random in-frame across entire scFv gene that were further recombined screened ribosome display. Knowledge potential insertion points from formed basis length sequence diversity novel insertional-scanning mutagenesis (ISM). An overall 256-fold improvement an anti-IL-13 antibody BAK1 as a result InDel combination known point mutations validates this approach suggests results conventional can synergize higher affinity.
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