Antibody biomarker discovery through in vitro directed evolution of consensus recognition epitopes

John T. Ballew, Joseph A Murray, Pekka Collin, Markku Mäki, Martin F. Kagnoff, Katri Kaukinen, Patrick S. Daugherty

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

To enable discovery of serum antibodies indicative of disease and simultaneously develop reagents suitable for diagnosis, in vitro directed evolution was applied to identify consensus peptides recognized by patients' serum antibodies. Bacterial cell-displayed peptide libraries were quantitatively screened for binders to serum antibodies from patients with celiac disease (CD), using cellsorting instrumentation to identify two distinct consensus epitope families specific to CD patients (PEQ and E/DxFVY/FQ). Evolution of the E/DxFVY/FQ consensus epitope identified a celiac-specific epitope, distinct from the two CD hallmark antigens tissue transglutaminase- 2 and deamidated gliadin, exhibiting 71% sensitivity and 99% specificity (n = 231). Expansion of the first-generation PEQ consensus epitope via in vitro evolution yielded octapeptides QPEQAFPE and PFPEQxFP that identified ω- and γ-gliadins, and their deamidated forms, as immunodominant B-cell epitopes in wheat and related cereal proteins. The evolved octapeptides, but not first-generation peptides, discriminated one-way blinded CD and non-CD sera (n = 78) with exceptional accuracy, yielding 100% sensitivity and 98% specificity. Because this method, termed antibody diagnostics via evolution of peptides, does not require prior knowledge of pathobiology, it may be broadly useful for de novo discovery of antibody biomarkers and reagents for their detection.

Original languageEnglish (US)
Pages (from-to)19330-19335
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume110
Issue number48
DOIs
StatePublished - Nov 26 2013

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Celiac Disease
Epitopes
Consensus
Biomarkers
Antibodies
Gliadin
Serum
Peptides
B-Lymphocyte Epitopes
Peptide Library
Sensitivity and Specificity
Immunodominant Epitopes
Abdomen
Triticum
Recognition (Psychology)
In Vitro Techniques
Antigens
Proteins
transglutaminase 2

ASJC Scopus subject areas

  • General

Cite this

Antibody biomarker discovery through in vitro directed evolution of consensus recognition epitopes. / Ballew, John T.; Murray, Joseph A; Collin, Pekka; Mäki, Markku; Kagnoff, Martin F.; Kaukinen, Katri; Daugherty, Patrick S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, No. 48, 26.11.2013, p. 19330-19335.

Research output: Contribution to journalArticle

Ballew, John T. ; Murray, Joseph A ; Collin, Pekka ; Mäki, Markku ; Kagnoff, Martin F. ; Kaukinen, Katri ; Daugherty, Patrick S. / Antibody biomarker discovery through in vitro directed evolution of consensus recognition epitopes. In: Proceedings of the National Academy of Sciences of the United States of America. 2013 ; Vol. 110, No. 48. pp. 19330-19335.
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