High-throughput method for ranking the affinity of peptide ligands selected from phage display libraries

A. González-Techera, M. Umpiérrez-Failache, S. Cardozo, G. Obal, O. Pritsch, Jerold A Last, S. J. Gee, B. D. Hammock, G. González-Sapienza

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The use of phage display peptide libraries allows rapid isolation of peptide ligands for any target selector molecule. However, due to differences in peptide expression and the heterogeneity of the phage preparations, there is no easy way to compare the binding properties of the selected clones, which operates as a major "bottleneck" of the technology. Here, we present the development of a new type of library that allows rapid comparison of the relative affinity of the selected peptides in a high-throughput screening format. As a model system, a phage display peptide library constructed on a phagemid vector that contains the bacterial alkaline phosphatase gene (BAP) was selected with an antiherbicide antibody. Due to the intrinsic switching capacity of the library, the selected peptides were transferred "en masse" from the phage coat protein to BAP. This was coupled to an optimized affinity ELISA where normalized amounts of the peptide - BAP fusion allow direct comparison of the binding properties of hundreds of peptide ligands. The system was validated by plasmon surface resonance experiments using synthetic peptides, showing that the method discriminates among the affinities of the peptides within 3 orders of magnitude. In addition, the peptide - BAP protein can find direct application as a tracer reagent.

Original languageEnglish (US)
Pages (from-to)993-1000
Number of pages8
JournalBioconjugate Chemistry
Volume19
Issue number5
DOIs
StatePublished - May 2008

Fingerprint

Bacteriophages
Peptides
Libraries
Display devices
Ligands
Throughput
Peptide Library
Alkaline Phosphatase
Phosphatases
Genes
Surface Plasmon Resonance
Gene Fusion
Capsid Proteins
Proteins
Surface plasmon resonance
Clone Cells
Enzyme-Linked Immunosorbent Assay
Screening
Antibodies
Fusion reactions

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

González-Techera, A., Umpiérrez-Failache, M., Cardozo, S., Obal, G., Pritsch, O., Last, J. A., ... González-Sapienza, G. (2008). High-throughput method for ranking the affinity of peptide ligands selected from phage display libraries. Bioconjugate Chemistry, 19(5), 993-1000. https://doi.org/10.1021/bc700279y

High-throughput method for ranking the affinity of peptide ligands selected from phage display libraries. / González-Techera, A.; Umpiérrez-Failache, M.; Cardozo, S.; Obal, G.; Pritsch, O.; Last, Jerold A; Gee, S. J.; Hammock, B. D.; González-Sapienza, G.

In: Bioconjugate Chemistry, Vol. 19, No. 5, 05.2008, p. 993-1000.

Research output: Contribution to journalArticle

González-Techera, A, Umpiérrez-Failache, M, Cardozo, S, Obal, G, Pritsch, O, Last, JA, Gee, SJ, Hammock, BD & González-Sapienza, G 2008, 'High-throughput method for ranking the affinity of peptide ligands selected from phage display libraries', Bioconjugate Chemistry, vol. 19, no. 5, pp. 993-1000. https://doi.org/10.1021/bc700279y
González-Techera, A. ; Umpiérrez-Failache, M. ; Cardozo, S. ; Obal, G. ; Pritsch, O. ; Last, Jerold A ; Gee, S. J. ; Hammock, B. D. ; González-Sapienza, G. / High-throughput method for ranking the affinity of peptide ligands selected from phage display libraries. In: Bioconjugate Chemistry. 2008 ; Vol. 19, No. 5. pp. 993-1000.
@article{4af6dc37f95c427ca358748bbf8a7b81,
title = "High-throughput method for ranking the affinity of peptide ligands selected from phage display libraries",
abstract = "The use of phage display peptide libraries allows rapid isolation of peptide ligands for any target selector molecule. However, due to differences in peptide expression and the heterogeneity of the phage preparations, there is no easy way to compare the binding properties of the selected clones, which operates as a major {"}bottleneck{"} of the technology. Here, we present the development of a new type of library that allows rapid comparison of the relative affinity of the selected peptides in a high-throughput screening format. As a model system, a phage display peptide library constructed on a phagemid vector that contains the bacterial alkaline phosphatase gene (BAP) was selected with an antiherbicide antibody. Due to the intrinsic switching capacity of the library, the selected peptides were transferred {"}en masse{"} from the phage coat protein to BAP. This was coupled to an optimized affinity ELISA where normalized amounts of the peptide - BAP fusion allow direct comparison of the binding properties of hundreds of peptide ligands. The system was validated by plasmon surface resonance experiments using synthetic peptides, showing that the method discriminates among the affinities of the peptides within 3 orders of magnitude. In addition, the peptide - BAP protein can find direct application as a tracer reagent.",
author = "A. Gonz{\'a}lez-Techera and M. Umpi{\'e}rrez-Failache and S. Cardozo and G. Obal and O. Pritsch and Last, {Jerold A} and Gee, {S. J.} and Hammock, {B. D.} and G. Gonz{\'a}lez-Sapienza",
year = "2008",
month = "5",
doi = "10.1021/bc700279y",
language = "English (US)",
volume = "19",
pages = "993--1000",
journal = "Bioconjugate Chemistry",
issn = "1043-1802",
publisher = "American Chemical Society",
number = "5",

}

TY - JOUR

T1 - High-throughput method for ranking the affinity of peptide ligands selected from phage display libraries

AU - González-Techera, A.

AU - Umpiérrez-Failache, M.

AU - Cardozo, S.

AU - Obal, G.

AU - Pritsch, O.

AU - Last, Jerold A

AU - Gee, S. J.

AU - Hammock, B. D.

AU - González-Sapienza, G.

PY - 2008/5

Y1 - 2008/5

N2 - The use of phage display peptide libraries allows rapid isolation of peptide ligands for any target selector molecule. However, due to differences in peptide expression and the heterogeneity of the phage preparations, there is no easy way to compare the binding properties of the selected clones, which operates as a major "bottleneck" of the technology. Here, we present the development of a new type of library that allows rapid comparison of the relative affinity of the selected peptides in a high-throughput screening format. As a model system, a phage display peptide library constructed on a phagemid vector that contains the bacterial alkaline phosphatase gene (BAP) was selected with an antiherbicide antibody. Due to the intrinsic switching capacity of the library, the selected peptides were transferred "en masse" from the phage coat protein to BAP. This was coupled to an optimized affinity ELISA where normalized amounts of the peptide - BAP fusion allow direct comparison of the binding properties of hundreds of peptide ligands. The system was validated by plasmon surface resonance experiments using synthetic peptides, showing that the method discriminates among the affinities of the peptides within 3 orders of magnitude. In addition, the peptide - BAP protein can find direct application as a tracer reagent.

AB - The use of phage display peptide libraries allows rapid isolation of peptide ligands for any target selector molecule. However, due to differences in peptide expression and the heterogeneity of the phage preparations, there is no easy way to compare the binding properties of the selected clones, which operates as a major "bottleneck" of the technology. Here, we present the development of a new type of library that allows rapid comparison of the relative affinity of the selected peptides in a high-throughput screening format. As a model system, a phage display peptide library constructed on a phagemid vector that contains the bacterial alkaline phosphatase gene (BAP) was selected with an antiherbicide antibody. Due to the intrinsic switching capacity of the library, the selected peptides were transferred "en masse" from the phage coat protein to BAP. This was coupled to an optimized affinity ELISA where normalized amounts of the peptide - BAP fusion allow direct comparison of the binding properties of hundreds of peptide ligands. The system was validated by plasmon surface resonance experiments using synthetic peptides, showing that the method discriminates among the affinities of the peptides within 3 orders of magnitude. In addition, the peptide - BAP protein can find direct application as a tracer reagent.

UR - http://www.scopus.com/inward/record.url?scp=44349139488&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=44349139488&partnerID=8YFLogxK

U2 - 10.1021/bc700279y

DO - 10.1021/bc700279y

M3 - Article

C2 - 18393454

AN - SCOPUS:44349139488

VL - 19

SP - 993

EP - 1000

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

SN - 1043-1802

IS - 5

ER -