Identification of IgE sequential epitopes of lentil (Len c 1) by means of peptide microarray immunoassay Andrea Vereda, MD, PhD, Doerthe A. Andreae,

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Identification of IgE sequential epitopes of lentil (Len c 1) by means of peptide microarray immunoassay Andrea Vereda, MD, PhD, Doerthe A. Andreae, MD, PhD, Jing Lin, PhD, Wayne G. Shreffler, MD, PhD, Maria Dolores Ibañez, MD, PhD, Javier Cuesta-Herranz, MD, PhD, Ludmilla Bardina, MSc, Hugh A. Sampson, MD Journal of Allergy and Clinical Immunology Volume 126, Issue 3, Pages 596-601.e1 (September 2010) DOI: 10.1016/j.jaci.2010.06.023 Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Average signal intensity (in digital fluorescence units [dfu]) plotted from the N-terminal to the C-terminal position of the Len c 1 sequence for patients (red) and negative control subjects (blue). Journal of Allergy and Clinical Immunology 2010 126, 596-601.e1DOI: (10.1016/j.jaci.2010.06.023) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Treeview display of hierarchic cluster analysis of IgE signal (positive in increasing brightness of red and negative in black). Peptides are in columns, and individual patients (arrays) are in rows. The arrows indicate peptide clusters (nodes) that were chosen for additional analysis of patients' characteristics. See the Methods and Results sections for details. Journal of Allergy and Clinical Immunology 2010 126, 596-601.e1DOI: (10.1016/j.jaci.2010.06.023) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Average signal intensity (in digital fluorescence units [dfu]) plotted from the N-terminal to the C-terminal position of the Len c 1 peptide for pooled sera preincubated with a single peptide for 2 hours (red) and pooled sera without preincubation (blue). Four separate experiments with different peptides were conducted (A-D). The arrows indicate the localization of the peptide used for the inhibition. Journal of Allergy and Clinical Immunology 2010 126, 596-601.e1DOI: (10.1016/j.jaci.2010.06.023) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Epitopes 1 and 4 as defined in this study superimposed on the 3-dimensional structure of the vicilin soy β-conglycin. Epitopes are located on the outside of the molecule. On top is epitope 1, and below is epitope 2. Journal of Allergy and Clinical Immunology 2010 126, 596-601.e1DOI: (10.1016/j.jaci.2010.06.023) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Sequence with highlighted epitopes. Journal of Allergy and Clinical Immunology 2010 126, 596-601.e1DOI: (10.1016/j.jaci.2010.06.023) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions