Development of peptide arrays for detection of IgE-binding epitopes in cow's milk allergens

Naoki Matsumoto, Mina Okochi, Miyoko Matsushima, Akiko Ogawa, Tomokazu Takase, Yasuko Yoshida, Mitsuo Kawase, Ken ichi Isobe, Tsutomu Kawabe, Hiroyuki Honda

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

Peptide arrays have become versatile tools for high throughput screening assays in biomedical and pharmaceutical research. In this study, we constructed a peptide array that contained linear peptide fragments reported as IgE-binding epitopes for cow's milk allergy (CMA). Various peptides with different solubility in aqueous solutions were dissolved in the buffer solutions containing sodium dodecyl sulfate, and we achieved a consistent spotting of peptide solutions using a piezoelectric ceramic micropump. The IgE-binding patterns were successfully detected by observing the binding of Alexa 647-labeled anti-human IgE using sera from CMA patients. Our technique in this study will provide a potent capability for the development of a peptide array for mapping IgE-epitopes in milk proteins, and it will help researchers better understand the IgE-epitopes associated with the clinical outcome of CMA.

Original languageEnglish
Pages (from-to)324-330
Number of pages7
JournalJournal of Bioscience and Bioengineering
Volume107
Issue number3
DOIs
Publication statusPublished - 2009 Mar

Keywords

  • Cow's milk allergy
  • IgE-binding epitopes
  • Peptide array
  • Piezoelectric ceramic micropump
  • spotting buffer

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