Skip Navigation



PEDS Advance Access published online on November 5, 2004
Protein Engineering Design and Selection, doi:10.1093/protein/gzh084
This Article
Right arrow FREE Full Text (PDF) Freely available
Right arrow Supplementary data
Right arrow All Versions of this Article:
17/10/731    most recent
gzh084v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Bessette, P. H.
Right arrow Articles by Daugherty, P. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bessette, P. H.
Right arrow Articles by Daugherty, P. S.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Received May 14, 2004
Revised October 2, 2004
Accepted October 23, 2004

Article

Rapid isolation of high affinity protein binding peptides using bacterial display

Paul H. Bessette 1, Jeffrey J. Rice 1, and Patrick S. Daugherty 1*

1 Department of Chemical Engineering, University of California, Santa Barbara, CA 93106, USA

* To whom correspondence should be addressed.
Patrick S. Daugherty, E-mail: psd{at}engineering.ucsb.edu


  Abstract

A robust bacterial display methodology has been developed enabling rapid isolation of peptides that bind to arbitrarily selected targets with high affinity. To demonstrate the utility of this approach, a large library (5x1010 clones) was constructed composed of random 15-mer peptide insertions constrained within a flexible, surface exposed loop of the E. coli outer membrane protein A (OmpA). The library was screened for binding to five unrelated proteins, including targets previously used in phage display selections: human serum albumin, anti-T7 epitope mAb, human C-reactive protein, HIV-1 GP120, and streptavidin. Two to four rounds of enrichment (2-4 days) were sufficient to enrich peptide ligands having high affinity for each of the target proteins. Strong amino acid consensus sequences were apparent for each of the targets tested, with up to seven consensus residues. Isolated peptide ligands remained functional when expressed as insertional fusions within a monomeric fluorescent protein. This bacterial display methodology thus provides an efficient process for identifying peptide affinity reagents and should be useful in a variety of molecular recognition applications.

Keywords: bacterial display; flow cytometry; library; peptide.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Protein Eng Des SelHome page
S. A. Kenrick and P. S. Daugherty
Bacterial display enables efficient and quantitative peptide affinity maturation
Protein Eng. Des. Sel., November 10, 2009; (2009) gzp065v1.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. D. Adams, U. Kim, and H. T. Soh
Multitarget magnetic activated cell sorter
PNAS, November 25, 2008; 105(47): 18165 - 18170.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
E. Schultheiss, S. Weiss, E. Winterer, R. Maas, E. Heinzle, and J. Jose
Esterase Autodisplay: Enzyme Engineering and Whole-Cell Activity Determination in Microplates with pH Sensors
Appl. Envir. Microbiol., August 1, 2008; 74(15): 4782 - 4791.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
M. Dai, J. Temirov, E. Pesavento, C. Kiss, N. Velappan, P. Pavlik, J.H. Werner, and A.R.M. Bradbury
Using T7 phage display to select GFP-based binders
Protein Eng. Des. Sel., July 1, 2008; 21(7): 413 - 424.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
J. J. Rice and P. S. Daugherty
Directed evolution of a biterminal bacterial display scaffold enhances the display of diverse peptides
Protein Eng. Des. Sel., July 1, 2008; 21(7): 435 - 442.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
N. Kronqvist, J. Lofblom, A. Jonsson, H. Wernerus, and S. Stahl
A novel affinity protein selection system based on staphylococcal cell surface display and flow cytometry
Protein Eng. Des. Sel., April 1, 2008; 21(4): 247 - 255.
[Abstract] [Full Text] [PDF]

Home page
 Microbiol. Mol. Biol. Rev.Home page
J. Jose and T. F. Meyer
The Autodisplay Story, from Discovery to Biotechnical and Biomedical Applications
Microbiol. Mol. Biol. Rev., December 1, 2007; 71(4): 600 - 619.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
R. Stork, D. Muller, and R. E. Kontermann
A novel tri-functional antibody fusion protein with improved pharmacokinetic properties generated by fusing a bispecific single-chain diabody with an albumin-binding domain from streptococcal protein G
Protein Eng. Des. Sel., November 3, 2007; (2007) gzm061v1.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
J. Lofblom, J. Sandberg, H. Wernerus, and S. Stahl
Evaluation of Staphylococcal Cell Surface Display and Flow Cytometry for Postselectional Characterization of Affinity Proteins in Combinatorial Protein Engineering Applications
Appl. Envir. Microbiol., November 1, 2007; 73(21): 6714 - 6721.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
C. Kiss, H. Fisher, E. Pesavento, M. Dai, R. Valero, M. Ovecka, R. Nolan, M. L. Phipps, N. Velappan, L. Chasteen, et al.
Antibody binding loop insertions as diversity elements
Nucleic Acids Res., November 14, 2006; 34(19): e132 - e132.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
K. T. Boulware and P. S. Daugherty
Protease specificity determination by using cellular libraries of peptide substrates (CLiPS)
PNAS, May 16, 2006; 103(20): 7583 - 7588.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
X. Hu, P. H. Bessette, J. Qian, C. D. Meinhart, P. S. Daugherty, and H. T. Soh
Marker-specific sorting of rare cells using dielectrophoresis
PNAS, November 1, 2005; 102(44): 15757 - 15761.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. E. Kohn and K. W. Plaxco
Engineering a signal transduction mechanism for protein-based biosensors
PNAS, August 2, 2005; 102(31): 10841 - 10845.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.