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

Anders Christiansen, Jens Vindahl Kringelum, Christian Skjødt Hansen, Katrine Lindholm Bøgh, Eric Sullivan, Jigar Patel, Neil M. Rigby, Thomas Eiwegger, Zsolt Szepfalusi, Federico De Masi, Morten Nielsen, Ole Lund, Martin Dufva

Research output: Contribution to journalJournal articleResearchpeer-review

424 Downloads (Pure)


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.
Original languageEnglish
Article number12913
JournalScientific Reports
Number of pages13
Publication statusPublished - 2015

Bibliographical note

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/


Dive into the research topics of 'High-throughput sequencing enhanced phage display enables the identification of patient-specific epitope motifs in serum'. Together they form a unique fingerprint.

Cite this