title: Ultrasensitive carbohydrate-peptide SPR imaging microarray for diagnosing IgE mediated peanut allergy.
abstract: Severity of peanut allergies is linked to allergen-specific immunoglobulin E (IgE) antibodies in blood, but diagnostics from assays using glycoprotein allergen mixtures may be inaccurate. Measuring IgEs specific to individual peptide and carbohydrate epitopes of allergenic proteins is promising. We report here the first immunoarray for IgEs utilizing both peptide and carbohydrate epitopes. A surface plasmon resonance imaging (SPRi) microarray was equipped with peptide and β-xylosyl glycoside (BXG) epitopes from major peanut allergen glycoprotein Arachis hypogaea h2 (Ara-h2). A monoclonal anti-IgE antibody was included as positive control. IgEs were precaptured onto magnetic beads loaded with polyclonal anti-IgE antibodies to enhance sensitivity and minimize non-specific binding. As little as 0.1 attomole (0.5 pg mL(-1)) IgE was detected from dilute serum in 45 min. IgEs binding to Ara-h2 peptide and BXG were quantified in 10 μL of patient serum and correlated with standard ImmunoCAP values.
authors.affiliation: Department of Chemistry, University of Connecticut, U-3060, Storrs, CT 06269-3060, USA. James.Rusling@uconn.edu.
authors.forename: Amit A
authors.initials: AA
authors.lastname: Joshi
authors.forename: Mark W
authors.initials: MW
authors.lastname: Peczuh
authors.forename: Challa V
authors.initials: CV
authors.lastname: Kumar
authors.forename: James F
authors.initials: JF
authors.lastname: Rusling
mesh_terms: D002241:Carbohydrates / Q000032:analysis*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D057230:Limit of Detection; D021183:Peanut Hypersensitivity / Q000175:diagnosis* / Q000276:immunology; D010455:Peptides / Q000032:analysis*; D020349:Surface Plasmon Resonance
chemical_list: D002241:Carbohydrates; D010455:Peptides; D007073:Immunoglobulin E