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Epitope‐Resolved Detection of Peanut‐Specific IgE Antibodies by Surface Plasmon Resonance Imaging |
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| Authors: | Min Shen Dr Amit A Joshi Dr Raghu Vannam Dr Chandra K Dixit Prof Dr Robert G Hamilton Prof Dr Challa V Kumar Prof Dr James F Rusling Prof Dr Mark W Peczuh |
| Affiliation: | 1. Department of Chemistry, University of Connecticut, Storrs, CT, USA;2. Division of Allergy and Clinical Immunology, Johns Hopkins University School of Medicine, Baltimore, MD, USA;3. Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT, USA;4. Department of Cell Biology, University of Connecticut Health Center, Farmington, CT, USA |
| Abstract: | Peanut allergy can be life‐threatening and is mediated by allergen‐specific immunoglobulin E (IgE) antibodies. Investigation of IgE antibody binding to allergenic epitopes can identify specific interactions underlying the allergic response. Here, we report a surface plasmon resonance imaging (SPRi) immunoassay for differentiating IgE antibodies by epitope‐resolved detection. IgE antibodies were first captured by magnetic beads bearing IgE ?‐chain‐specific antibodies and then introduced into an SPRi array immobilized with epitopes from the major peanut allergen glycoprotein Arachis hypogaea h2 (Ara h2). Differential epitope responses were achieved by establishing a binding environment that minimized cross‐reactivity while maximizing analytical sensitivity. IgE antibody binding to each Ara h2 epitope was distinguished and quantified from patient serum samples (10 μL each) in a 45 min assay. Excellent correlation of Ara h2‐specific IgE values was found between ImmunoCAP assays and the new SPRi method. |
| Keywords: | immunoassays immunoglobulin E magnetic bead peanut allergy surface plasmon resonance (SPR) imaging |