Abstract
IgE epitope mapping has the potential to become an additional tool for food allergy diagnosis/prognosis and to lead to a better understanding of the pathogenesis and tolerance induction of food allergy. Due to its ability to screen thousands of targets in parallel using small volumes of sample, peptide microarray has greatly facilitated large-scale IgE epitope mapping. In the past 10 years, we have developed and optimized a reliable and sensitive peptide microarray immunoassay, which has been successfully applied for IgE epitope mapping of many food allergens in our lab. Here, we describe the method of performing the peptide microarray immunoassay for IgE epitope mapping. In addition, we have upgraded the microarray platform to measure antibody affinity by adding one additional competition step, which is also described in this chapter.
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References
Sicherer SH, Sampson HA (2014) Food allergy: epidemiology, pathogenesis, diagnosis, and treatment. J Allergy Clin Immunol 133(2):291–307
Lin J, Sampson HA (2009) The role of immunoglobulin E-binding epitopes in the characterization of food allergy. Curr Opin Allergy Clin Immunol 9(4):357–363
Shreffler WG, Lencer DA, Bardina L et al (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(4):893–899
Lin J, Bardina L, Shreffler WG et al (2009) Development of a novel peptide microarray for large-scale epitope mapping of food allergens. J Allergy Clin Immunol 124(2):315–322
Lin J, Bruni FM, Fu ZY et al (2012) A bioinformatics approach to identify patients with symptomatic peanut allergy using peptide microarray immunoassay. J Allergy Clin Immunol 129(5):1321–1328
Ayuso R, Sanchez-Garcia S, Lin J et al (2010) Greater epitope recognition of shrimp allergens by children than by adults suggests that shrimp sensitization decreases with age. J Allergy Clin Immunol 125(6):1286–1293
Flinterman AE, Knol EF, Lencer DA et al (2008) Peanut epitopes for IgE and IgG4 in peanut-sensitized children in relation to severity of peanut allergy. J Allergy Clin Immunol 121(3):737–743
Wang J, Lin J, Bardina L et al (2010) Correlation of IgE/IgG4 milk epitopes and affinity of milk-specific IgE antibodies with different phenotypes of clinical milk allergy. J Allergy Clin Immunol 125(3):695–702
Han Y, Lin J, Bardina L et al (2016) What characteristics confer proteins the ability to induce allergic responses? IgE epitope mapping and comparison of the structure of soybean 2S albumins and Ara h 2. Molecules 21(5):622
McHeyzer-Williams MG, McHeyzer-Williams LJ (2005) Antigen-specific memory B cell development. Annu Rev Immunol 23:487–513
Christensen LH, Holm J, Lund G et al (2008) Several distinct properties of the IgE repertoire determine effector cell degranulation in response to allergen challenge. J Allergy Clin Immunol 122(2):298–304
El-Khouly F, Lewis SA, Pons L et al (2007) IgG and IgE avidity characteristics of peanut allergic individuals. Pediatr Allergy Immunol 18:607–613
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Lin, J., Sampson, H.A. (2017). IgE Epitope Mapping Using Peptide Microarray Immunoassay. In: Lin, J., Alcocer, M. (eds) Food Allergens. Methods in Molecular Biology, vol 1592. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6925-8_14
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DOI: https://doi.org/10.1007/978-1-4939-6925-8_14
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6923-4
Online ISBN: 978-1-4939-6925-8
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