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In vitro evolution of antibody affinity via insertional scanning mutagenesis of an entire antibody variable region

2020; National Academy of Sciences; Volume: 117; Issue: 44 Linguagem: Inglês

10.1073/pnas.2002954117

1091-6490

Kalliopi Skamaki, Stéphane Emond, Matthieu Chodorge, John L. Andrews, D. Gareth Rees, Daniel A. Cannon, Bojana Popovic, Andrew Buchanan, Ralph Minter, Florian Hollfelder,

Glycosylation and Glycoproteins Research

Significance Insertion/deletion (InDel) mutations play key roles in genome and protein evolution. Despite their prominence in evolutionary history, the potential of InDels for changing function in protein engineering by directed evolution remains unexplored. Instead point mutagenesis is widely used. Here we create antibody libraries containing InDels and demonstrate that affinity maturation can be achieved in this way, establishing an alternative to the point mutation strategies employed in all previous in vitro selections. These InDels mirror the observation of considerable length variation in loops of natural antibodies originating from the same germline genes and be combined with point mutations, making both natural sources of functional innovation available for artificial evolution in the test tube.