Allergen micro-bead array for IgE detection: A feasibility study using allergenic molecules tested on a flexible multiplex flow cytometric immunoassay

Debora Pomponi, Maria Livia Bernardi, Marina Liso, Paola Palazzo, Lisa Tuppo, Chiara Rafaiani, Mario Santoro, Alexis Labrada, Maria Antonietta Ciardiello, Adriano Mari, Enrico Scala

Research output: Contribution to journalArticle

Abstract

Background: Allergies represent the most prevalent non infective diseases worldwide. Approaching IgE-mediated sensitizations improved much by adopting allergenic molecules instead of extracts, and by using the micro-technology for multiplex testing. Objective and Methods: To provide a proof-of-concept that a flow cytometric bead array is a feasible mean for the detection of specific IgE reactivity to allergenic molecules in a multiplex-like way. A flow cytometry Allergenic Molecule-based micro-bead Array system (ABA) was set by coupling allergenic molecules with commercially available micro-beads. Allergen specific polyclonal and monoclonal antibodies, as well as samples from 167 allergic patients, characterized by means of the ISAC microarray system, were used as means to show the feasibility of the ABA. Three hundred and thirty-six sera were tested for 1 or more of the 16 selected allergens, for a total number of 1,519 tests on each of the two systems. Results: Successful coupling was initially verified by detecting the binding of rabbit polyclonal IgG, mouse monoclonal, and pooled human IgE toward three allergens, namely nDer s 1, nPen m 1, and nPru p 3. The ABA assay showed to detect IgE to nAct d 1, nAct d 11, rAln g 1, nAmb a 1, nArt v 3, rBet v 1, rCor a 1, nCup a 1, nDer p 1, nDer s 1, rHev b 5, nOle e 1, rPar j 2, nPen m 1, rPhl p 1, and nPru p 3. Results obtained by ABA IgE testing were highly correlated to ISAC testing (r = 0.87, p

Original languageEnglish
Article numbere35697
JournalPLoS One
Volume7
Issue number4
DOIs
Publication statusPublished - Apr 17 2012

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

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    Pomponi, D., Bernardi, M. L., Liso, M., Palazzo, P., Tuppo, L., Rafaiani, C., Santoro, M., Labrada, A., Ciardiello, M. A., Mari, A., & Scala, E. (2012). Allergen micro-bead array for IgE detection: A feasibility study using allergenic molecules tested on a flexible multiplex flow cytometric immunoassay. PLoS One, 7(4), [e35697]. https://doi.org/10.1371/journal.pone.0035697