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Protein evolution with an expanded genetic code.
- Source :
-
Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2008 Nov 18; Vol. 105 (46), pp. 17688-93. Date of Electronic Publication: 2008 Nov 11. - Publication Year :
- 2008
-
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.
- Subjects :
- Amino Acids genetics
Antibodies, Viral immunology
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Genetic Vectors
Germ Cells immunology
HIV immunology
HIV Envelope Protein gp120 immunology
Humans
Peptide Library
Sequence Analysis, Protein
Tyrosine analogs & derivatives
Tyrosine metabolism
Directed Molecular Evolution
Genetic Code
Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1091-6490
- Volume :
- 105
- Issue :
- 46
- Database :
- MEDLINE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Publication Type :
- Academic Journal
- Accession number :
- 19004806
- Full Text :
- https://doi.org/10.1073/pnas.0809543105