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IgE and IgG4 Epitope Mapping of Food Allergens with a Peptide Microarray Immunoassay

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Peptide Microarrays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2578))

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Abstract

Peptide microarrays are a powerful tool to identify linear epitopes of food allergens in a high-throughput manner. The main advantages of the microarray-based immunoassay are as follows: the possibility to assay thousands of targets simultaneously, the requirement of a low volume of serum, the more robust statistical analysis, and the possibility to test simultaneously several immunoglobulin subclasses. Among them, the last one has a special interest in the field of food allergy, because the development of tolerance to food allergens has been associated with a decrease in IgE and an increase in IgG4 levels against linear epitopes. However, the main limitation to the clinical use of microarray is the automated analysis of the data. Recent studies mapping the linear epitopes of food allergens with peptide microarray immunoassays have identified peptide biomarkers that can be used for early diagnosis of food allergies and to predict their severity or the self-development of tolerance. Using this approach, we have worked on epitope mapping of the two most important food allergens in the Spanish population, cow’s milk, and chicken eggs. The final aim of these studies is to define subsets of peptides that could be used as biomarkers to improve the diagnosis and prognosis of food allergies. This chapter describes the protocol to produce microarrays using a library of overlapping peptides corresponding to the primary sequences of food allergens and data acquisition and analysis of IgE and IgG4 binding epitopes.

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Acknowledgments

We wish to thank Nuria Corcoles for excellent technical assistance. We also thank Raquel Rodriguez and Joon Won Park for their help and revision of the illustration of dendron-modified slides. This work was supported by grants from the Fondo de Investigación Sanitaria-Instituto de Salud Carlos III and the Sociedad Española de Alergología e Inmunología Clínica.

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Authors and Affiliations

  1. Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal - Unidad de Microarray IRYCIS, Madrid, Spain

    Javier Martínez-Botas & Carlos Fernández-Lozano

  2. CIBER de Fisiopatología de la Obesidad y Nutrición (CIBEROBN) - Instituto de la Salud Carlos III, Madrid, Spain

    Javier Martínez-Botas

  3. Servicio de Alergología, Hospital Universitario Ramón y Cajal - Unidad de Microarray IRYCIS, Madrid, Spain

    Aida Vaquero-Rey & Belén de la Hoz

  4. Red de Alergia ARADyAL (RD16/0006/0009) - Instituto de la Salud Carlos III, Madrid, Spain

    Belén de la Hoz

Authors
  1. Javier Martínez-Botas
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  2. Carlos Fernández-Lozano
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  3. Aida Vaquero-Rey
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  4. Belén de la Hoz
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Editor information

Editors and Affiliations

  1. Istituto di Scienze e Tecnologie Chimiche (SCITEC), Consiglio Nazionale delle Ricerche, Milano, Italy

    Marina Cretich

  2. Istituto di Scienze e Tecnologie Chimiche (SCITEC), Consiglio Nazionale delle Ricerche, Milano, Italy

    Alessandro Gori

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Martínez-Botas, J., Fernández-Lozano, C., Vaquero-Rey, A., de la Hoz, B. (2023). IgE and IgG4 Epitope Mapping of Food Allergens with a Peptide Microarray Immunoassay. In: Cretich, M., Gori, A. (eds) Peptide Microarrays. Methods in Molecular Biology, vol 2578. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2732-7_16

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  • DOI: https://doi.org/10.1007/978-1-0716-2732-7_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2731-0

  • Online ISBN: 978-1-0716-2732-7

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