Protein evolution with an expanded genetic code

Chang C. Liu; Antha V. Mack; Meng Lin Tsao; Jeremy H. Mills; Soo Lee Hyun; Hyeryun Choe; Michael Farzan; Peter G. Schultz; Vaughn V. Smider

doi:10.1073/pnas.0809543105

Protein evolution with an expanded genetic code

Chang C. Liu, Antha V. Mack, Meng Lin Tsao, Jeremy H. Mills, Soo Lee Hyun, Hyeryun Choe, Michael Farzan, Peter G. Schultz, Vaughn V. Smider

Research output: Contribution to journal › Article › peer-review

120 Scopus citations

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.

Original language	English (US)
Pages (from-to)	17688-17693
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	105
Issue number	46
DOIs	https://doi.org/10.1073/pnas.0809543105
State	Published - Nov 18 2008
Externally published	Yes

Keywords

Directed evolution
Phage display
Unnatural amino acids

ASJC Scopus subject areas

General

Access to Document

10.1073/pnas.0809543105

Cite this

@article{e78a489e6fe2425681bce7b7517fc91b,

title = "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 na{\"i}ve germline scFv antibody library in which six residues in VH 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.",

keywords = "Directed evolution, Phage display, Unnatural amino acids",

author = "Liu, {Chang C.} and Mack, {Antha V.} and Tsao, {Meng Lin} and Mills, {Jeremy H.} and Hyun, {Soo Lee} and Hyeryun Choe and Michael Farzan and Schultz, {Peter G.} and Smider, {Vaughn V.}",

year = "2008",

month = nov,

day = "18",

doi = "10.1073/pnas.0809543105",

language = "English (US)",

volume = "105",

pages = "17688--17693",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "46",

}

TY - JOUR

T1 - Protein evolution with an expanded genetic code

AU - Liu, Chang C.

AU - Mack, Antha V.

AU - Tsao, Meng Lin

AU - Mills, Jeremy H.

AU - Hyun, Soo Lee

AU - Choe, Hyeryun

AU - Farzan, Michael

AU - Schultz, Peter G.

AU - Smider, Vaughn V.

PY - 2008/11/18

Y1 - 2008/11/18

N2 - 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 VH 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.

AB - 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 VH 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.

KW - Directed evolution

KW - Phage display

KW - Unnatural amino acids

UR - http://www.scopus.com/inward/record.url?scp=56649124785&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=56649124785&partnerID=8YFLogxK

U2 - 10.1073/pnas.0809543105

DO - 10.1073/pnas.0809543105

M3 - Article

C2 - 19004806

AN - SCOPUS:56649124785

SN - 0027-8424

VL - 105

SP - 17688

EP - 17693

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 46

ER -

Protein evolution with an expanded genetic code

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this