文章基本信息

标题：In vitro evolution of antibody affinity via insertional scanning mutagenesis of an entire antibody variable region
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作者：Kalliopi Skamaki ; Stephane Emond ; Matthieu Chodorge 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2020
卷号：117
期号：44
页码：27307-27318
DOI：10.1073/pnas.2002954117
出版社：The National Academy of Sciences of the United States of America
摘要：We report a systematic combinatorial exploration of affinity enhancement of antibodies by insertions and deletions (InDels). Transposon-based introduction of InDels via the method TRIAD (transposition-based random insertion and deletion mutagenesis) was used to generate large libraries with random in-frame InDels across the entire single-chain variable fragment 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 (InScaM). 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 mutagenesis and conventional exploration of point mutations can synergize to generate antibodies with higher affinity.
关键词：antibody ; directed evolution ; InDel ; protein engineering