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Epitope Mapping Using Peptide Microarray in Autoantibody Profiling

: Henkel, S.; Wellhausen, R.; Woitalla, D.; Marcus, K.; May, C.


Li, P.H.C.:
Microarray technology. Methods and applications
New York: Humana Press, 2015 (Methods in molecular biology 1368)
ISBN: 978-1-4939-3135-4
Book Article
Fraunhofer IZI ()

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.