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.
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Acknowledgments
This work was supported by P.U.R.E. (Protein Unit for Research in Europe), which is a project of the federal state of Nordrhein-Westfalen, Germany, and by the Ministerium für Innovation, Wissenschaft und Forschung des Landes Nordrhein-Westfalen and WTZ Brasilien, which is a project of the German Ministry of Education and Research (BMBF), Germany (01DN14023).
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Henkel, S., Wellhausen, R., Woitalla, D., Marcus, K., May, C. (2016). Epitope Mapping Using Peptide Microarray in Autoantibody Profiling. In: Li, P., Sedighi, A., Wang, L. (eds) Microarray Technology. Methods in Molecular Biology, vol 1368. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3136-1_15
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DOI: https://doi.org/10.1007/978-1-4939-3136-1_15
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