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Protein evolution with an expanded genetic code.


ABSTRACT: We have devised a phage display system in which an expanded genetic code is available for directed evolution. This system allows selection to yield proteins containing unnatural amino acids should such sequences functionally outperform ones containing only the 20 canonical amino acids. We have optimized this system for use with several unnatural amino acids and provide a demonstration of its utility through the selection of anti-gp120 antibodies. One such phage-displayed antibody, selected from a naïve germline scFv antibody library in which six residues in V(H) CDR3 were randomized, contains sulfotyrosine and binds gp120 more effectively than a similarly displayed known sulfated antibody isolated from human serum. These experiments suggest that an expanded "synthetic" genetic code can confer a selective advantage in the directed evolution of proteins with specific properties.

SUBMITTER: Liu CC 

PROVIDER: S-EPMC2584696 | biostudies-literature | 2008 Nov

REPOSITORIES: biostudies-literature

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Protein evolution with an expanded genetic code.

Liu Chang C CC   Mack Antha V AV   Tsao Meng-Lin ML   Mills Jeremy H JH   Lee Hyun Soo HS   Choe Hyeryun H   Farzan Michael M   Schultz Peter G PG   Smider Vaughn V VV  

Proceedings of the National Academy of Sciences of the United States of America 20081111 46


We have devised a phage display system in which an expanded genetic code is available for directed evolution. This system allows selection to yield proteins containing unnatural amino acids should such sequences functionally outperform ones containing only the 20 canonical amino acids. We have optimized this system for use with several unnatural amino acids and provide a demonstration of its utility through the selection of anti-gp120 antibodies. One such phage-displayed antibody, selected from  ...[more]

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