Summary
With the evolution of peptide synthesis techniques and microarray technology, it is now possible to map and characterize allergenic epitopes on a microarray platform. The peptide microarray-based immunoassay allows simultaneous analysis of thousands of target peptides using small volumes of diluted serum samples, and has a promising future to become a superior testing tool for aspects of food allergy diagnosis and prognosis, as well as for designing recombinant allergens for safe immunotherapy. Characterization of allergenic epitopes provides fundamental knowledge for understanding mechanisms of food allergy. This chapter describes in detail the development of a sensitive and reliable peptide microarray-based immunoassay. The information includes a comparison of different slide substrates, effect of buffers on spot morphology, performance of printing pins, immunolabeling detection systems with different levels of sensitivity, and suggested approaches to data analysis.
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References
Sampson, H. A. (2005) Food allergy–accurately identifying clinical reactivity. Allergy 60, 19–24.
Sampson, H. A. (2001) Utility of food-specific IgE concentrations in predicting symptomatic food allergy. J. Allergy Clin. Immunol. 107, 891–896.
Chatchatee, P., Järvinen, K. M., Bardina, L., Vila, L., Beyer, K., and Sampson, H. A. (2001) Identification of IgE and IgG binding epitopes on beta- and kappa-casein in cow’s milk allergic patients. Clin. Exp. Allergy 31, 1256–1262.
Jarvinen, K. M., Beyer, K., Vila, L., Chatchatee, P., Busse, P. J., and Sampson, H. A. (2002) B-cell epitopes as a screening instrument for persistent cow’s milk allergy. J. Allergy Clin. Immunol. 110, 293–297.
Beyer, K., Ellman-Grunther, L., Järvinen, K. M., Wood, R. A., Hourihane, J., and Sampson, H. A. (2003) Measurement of peptide-specific IgE as an additional tool in identifying patients with clinical reactivity to peanuts. J. Allergy Clin. Immunol. 112, 202–207.
Cooke, S. K. and Sampson, H. A. (1997) Allergenic properties of ovomucoid in man. J. Immunol. 159, 2026–2032.
Bannon, G. A., Cockrell, G., Connaughton, C., West, C. M., Helm, R., Stanley, J. S., King, N., Rabjohn, P., Sampson, H. A., and Burks, A. W. (2001) Engineering, characterization and in vitro efficacy of the major peanut allergens for use in immunotherapy. Int. Arch. Allergy Imm. 124, 70–72.
Li, X. M., Srivastava, K., Grishin, A., Huang, C. K., Schofield, B., Burks, W., and Sampson, H. A. (2003) Persistent protective effect of heat-killed Escherichia coli producing “engineered,” recombinant peanut proteins in a murine model of peanut allergy. J. Allergy Clin. Immunol. 112, 159–167.
Valenta, R. and Niederberger, V. (2007) Recombinant allergens for immunotherapy. J. Allergy Clin. Immunol. 119, 826–830.
Frank, R. (2002) The SPOT synthesis technique–Synthetic peptide arrays on membrane supports–principles and applications. J. Immunol. Methods 267, 13–26.
Hiller, R., Laffer, S., Harwanegg, C., Huber, M., Schmidt, W. M., Twardosz, A., Barletta, B., Becker, W. M., Blaser, K., Breiteneder, H., Chapman, M., Crameri, R., Duchêne, M., Ferreira, F., Fiebig, H., Hoffmann-Sommergruber, K., King, T. P., Kleber-Janke, T., Kurup, V. P., Lehrer, S. B., Lidholm, J., Müller, U., Pini, C., Reese, G., Scheiner, O., Scheynius, A., Shen, H. D., Spitzauer, S., Suck, R., Swoboda, I., Thomas, W., Tinghino, R., Van Hage-Hamsten, M., Virtanen, T., Kraft, D., Müller, M. W., and Valenta, R. (2002) Microarrayed allergen molecules: diagnostic gatekeepers for allergy treatment. FASEB J. 16, 414–416.
Harwanegg, C., Laffer, S., Hiller, R., Mueller, M. W., Kraft, D., Spitzauer, S., and Valenta, R. (2003) Microarrayed recombinant allergens for diagnosis of allergy. Clin. Exp. Allergy 33, 7–13.
MacBeath, G. and Schreiber, S. L. (2000) Printing proteins as microarrays for high-throughput function determination. Science 289, 1760–1763.
Shin, D. S., Kim, D. H., Chung, W. J., and Lee, Y. S. (2005) Combinatorial solid phase peptide synthesis and bioassays. J. Biochem. Mol. Biol. 38, 517–525.
Panicker, R. C., Huang, X., and Yao, S. Q. (2004) Recent advances in peptide-based microarray technologies. Comb. Chem. High Throughput Screen. 7, 547–556.
Shreffler, W. G., Beyer, K., Chu, T. H., Burks, A. W., and Sampson, H. A. (2004) Microarray immunoassay: Association of clinical history, in vitro IgE function, and heterogeneity of allergenic peanut epitopes. J. Allergy Clin. Immunol. 113, 776–782.
Shreffler, W. G., Lencer, D. A., Bardina, L., and Sampson, H. A. (2005) IgE and IgG(4) epitope mapping by microarray immunoassay reveals the diversity of immune response to the peanut allergen, Ara h 2. J. Allergy Clin. Immunol. 116, 893–899.
Lencer, D. A., Rosenfeld, L., Bardina, L., Shreffler, W. G., Sampson, H. A., and Beyer, K. (2006) Determination of allergenic cross-reactivity between 11S-globulins (seed storage proteins) in peanut, tree-nut and sesame allergic patients using peptide microarray immunoassay. J. Allergy Clin. Immunol.117, S34.
Wang, J., Bardina, L., Lencer, D., Shreffler, W. G., and Sampson, H. A. (2006) Determination of epitope diversity in cow’s milk hypersensitive using microarray immunassay. J. Allergy Clin. Immunol. 117, S39.
Kusnezow, W. and Hoheisel, J. D. (2003) Solid supports for microarray immunoassays. J. Mol. Recognit. 16, 165–176.
Acknowledgements
The authors acknowledge the contribution of Yongchao Ge, Ph.D., Assistant Professor Mount Sinai, Department of Neurology, to the statistical analysis methods and authoring of an R analysis script used for some analysis steps. We also thank Todd Martinsky, Telechem International Inc. for technical advice and support.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Lin, J., Bardina, L., Shreffler, W.G. (2009). Microarrayed Allergen Molecules for Diagnostics of Allergy. In: Schutkowski, M., Reineke, U. (eds) Epitope Mapping Protocols. Methods in Molecular Biology™, vol 524. Humana Press. https://doi.org/10.1007/978-1-59745-450-6_19
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DOI: https://doi.org/10.1007/978-1-59745-450-6_19
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