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Cell Surface Display Yields Evolvable, Clickable Antibody Fragments

Van Deventer, James A. and Yuet, Kai P. and Yoo, Tae Hyeon and Tirrell, David A. (2014) Cell Surface Display Yields Evolvable, Clickable Antibody Fragments. ChemBioChem, 15 (12). pp. 1777-1781. ISSN 1439-4227. PMCID PMC4199383. doi:10.1002/cbic.201402184. https://resolver.caltech.edu/CaltechAUTHORS:20140728-141903775

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Abstract

Non-canonical amino acids (ncAAs) provide powerful tools for engineering the chemical and physical properties of proteins. However, introducing ncAAs into proteins can affect protein properties in unpredictable ways, thus necessitating screening efforts to identify mutants with desirable properties. In this work, we describe an Escherichia coli cell surface display platform for the directed evolution of clickable antibody fragments. This platform enabled isolation of antibody fragments with improved digoxigenin binding and modest affinity maturation in several different ncAA contexts. Azide-functionalized fragments exhibited improved binding kinetics relative to their methionine counterparts, facile chemical modification through azide–alkyne cycloaddition, and retention of binding properties after modification. The results described here suggest new possibilities for protein engineering, including modulation of molecular recognition events by ncAAs and direct screening of libraries of chemically modified proteins.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1002/cbic.201402184DOIArticle
http://onlinelibrary.wiley.com/doi/10.1002/cbic.201402184/suppinfoPublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4199383/ADSArticle
ORCID:
AuthorORCID
Yuet, Kai P.0000-0002-1381-8923
Yoo, Tae Hyeon0000-0003-1448-3165
Tirrell, David A.0000-0003-3175-4596
Additional Information:© 2014 Wiley-VCH Verlag GmbH & Co. Article first published online: 17 Jul 2014. We thank Mona Shahgholi for MALDI mass spectrometry assistance at the Caltech Chemistry Mass Spectrometry facility, Igor Antoshechkin and the Millard and Muriel Jacobs Genetics and Genomics Laboratory for assistance with high-throughput sequencing, Cynthia Shuman of GE Healthcare for Biacore kinetic assay guidance, Janek Szychowski and Alborz Mahdavi for materials, and Maren Buck for helpful comments on the manuscript. This work was supported by NIH grant R01 GM62523. J.A.V. was supported in part by a National Defense Science and Engineering Graduate (NDSEG) fellowship, and K.P.Y. in part by a National Science Foundation (NSF) Graduate Fellowship.
Funders:
Funding AgencyGrant Number
NIHR01 GM62523
National Defense Science and Engineering Graduate (NDSEG) FellowshipUNSPECIFIED
NSF Graduate Research FellowshipUNSPECIFIED
Subject Keywords:antibodies; click chemistry; directed evolution; high-throughput screening; non-canonical amino acids
Issue or Number:12
PubMed Central ID:PMC4199383
DOI:10.1002/cbic.201402184
Record Number:CaltechAUTHORS:20140728-141903775
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20140728-141903775
Official Citation:Van Deventer, J. A., Yuet, K. P., Yoo, T. H. and Tirrell, D. A. (2014), Cell Surface Display Yields Evolvable, Clickable Antibody Fragments. ChemBioChem, 15: 1777–1781. doi: 10.1002/cbic.201402184
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:47532
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:28 Jul 2014 21:42
Last Modified:10 Nov 2021 17:47

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