title: Epitope Mapping Using Peptide Microarray in Autoantibody Profiling.
abstract: The use of peptide microarrays for epitope mapping of autoantibodies greatly facilitates the early diagnosis of allergic, cytotoxin-associated diseases and especially inflammatory diseases. A common approach to create the microarrays utilizes nitrocellulose-coated glass slides for peptide probe binding, which is based on surface adsorption. Advantages of this method include excellent peptide binding capacity and long-term stability. To ensure equal accessibility to all antibodies on the peptide microarray during epitope mapping, all probes are immobilized in a random manner, thus avoiding concentration-dependent effects on signal intensity.In this chapter, we provide a step-by-step protocol on how to construct the peptide microarrays and perform epitope mapping of autoantibodies using them. Finally we present a comparative approach for the evaluation of the data.
authors.affiliation: Medizinisches Proteom-Center, Ruhr-Universität Bochum, Universitätsstr. 150, 44801, Bochum, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Henkel
authors.affiliation: Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytics and Bioprocesses (IZI-BB), Am Mühlenberg 13, 14476, Potsdam-Golm, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Wellhausen
authors.affiliation: St. Josef Hospital Bochum, Gudrunstraße 56, 44791, Bochum, Germany.|St. Josef Hospital Kupferdreh, Heidbergweg 22-24, 45257, Essen, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Woitalla
authors.affiliation: Medizinisches Proteom-Center, Ruhr-Universität Bochum, Universitätsstr. 150, 44801, Bochum, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Marcus
authors.affiliation: Medizinisches Proteom-Center, Ruhr-Universität Bochum, Universitätsstr. 150, 44801, Bochum, Germany. Caroline.May@rub.de.
authors.forename: Caroline
authors.initials: C
authors.lastname: May
mesh_terms: D001323:Autoantibodies / Q000097:blood / Q000737:chemistry / Q000276:immunology*; D001800:Blood Specimen Collection; D018604:Epitope Mapping / Q000379:methods*; D006801:Humans; D010455:Peptides / Q000097:blood; D040081:Protein Array Analysis / Q000379:methods*
chemical_list: D001323:Autoantibodies; D010455:Peptides
keywords: Autoantibody profiling; Epitope mapping; Nitrocellulose-coated glass slides; Peptide microarray