High-throughput sequencing enhanced phage display enables the identification of patient-specific epitope motifs in serum

Phage display is a prominent screening technique with a multitude of applications including therapeutic antibody development and mapping of antigen epitopes. In this study, phages were selected based on their interaction with patient serum and exhaustively characterised by high-throughput sequencing...

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Bibliographic Details
Published in:Scientific reports 2015-08, Vol.5 (1), p.12913-12913, Article 12913
Main Authors: Christiansen, Anders, Kringelum, Jens V., Hansen, Christian S., Bøgh, Katrine L., Sullivan, Eric, Patel, Jigar, Rigby, Neil M., Eiwegger, Thomas, Szépfalusi, Zsolt, Masi, Federico de, Nielsen, Morten, Lund, Ole, Dufva, Martin
Format: Article
Language:English
Subjects:
38/1
38/47
38/79
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631/1647/2163
631/1647/514/2254
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Allergens
Allergies
Amino Acid Motifs
Ara h 1 antigen
Bioinformatics
Biological samples
Epitope mapping
Epitopes - blood
Epitopes - genetics
Female
Food allergies
Gene Library
High-Throughput Nucleotide Sequencing
Humanities and Social Sciences
Humans
Male
multidisciplinary
Next-generation sequencing
Peanut Hypersensitivity - blood
Peanut Hypersensitivity - genetics
Peanuts
Peptides
Phage display
Phages
Protein Array Analysis
Science
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Summary:Phage display is a prominent screening technique with a multitude of applications including therapeutic antibody development and mapping of antigen epitopes. In this study, phages were selected based on their interaction with patient serum and exhaustively characterised by high-throughput sequencing. A bioinformatics approach was developed in order to identify peptide motifs of interest based on clustering and contrasting to control samples. Comparison of patient and control samples confirmed a major issue in phage display, namely the selection of unspecific peptides. The potential of the bioinformatic approach was demonstrated by identifying epitopes of a prominent peanut allergen, Ara h 1, in sera from patients with severe peanut allergy. The identified epitopes were confirmed by high-density peptide micro-arrays. The present study demonstrates that high-throughput sequencing can empower phage display by (i) enabling the analysis of complex biological samples, (ii) circumventing the traditional laborious picking and functional testing of individual phage clones and (iii) reducing the number of selection rounds.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep12913