Abstract
Protein microarray is a miniaturized multi-analyte, solid-phased immunoassay where thousands of immobilized individual protein spots on a microscopic slide bind are bound to specific antibodies (immunoglobulins) from serum samples, which are then detected by fluorescent labeling. The image processing and pattern recognition are then quantitatively analyzed using advanced algorithms. Here, we describe the use of an in-house-produced complex protein microarray containing extracts and pure proteins that has been probed with antibodies present in the horse sera and detection by fluorophore-conjugated antibody and data analysis. The flexibility of the number and types of proteins that can be printed on the microarray allows different set of specific IgE immunoassay analysis to be carried out.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Renault NK, Gaddipati SR, Wulfert F et al (2011) Multiple protein extract microarray for profiling human food-specific immunoglobulins A, M, G and E. J Immunol Methods 364(1–2):21–32
Renault NK, Mirotti L, Alcocer MJ (2007) Biotechnologies in new high-throughput food allergy tests: why we need them. Biotechnol Lett 29(3):333–339
Wulfert F, Sanyasi G, Tongen L et al (2012) Prediction of tolerance in children with IgE mediated cow’s milk allergy by microarray profiling and chemometric approach. J Immunol Methods 382(1–2):48–57
Marti E, Wang X, Jambari NN et al (2015) Novel in vitro diagnosis of equine allergies using a protein array and mathematical modelling approach: a proof of concept using insect bite hypersensitivity. Vet Immunol Immunopathol 167(3–4):171–177
Eriksson L, Trygg J, Wold S (2014) A chemometrics toolbox based on projections and latent variables. J Chemometr 28(5):332–346
Wold S, Sjostrom M, Eriksson L (2001) PLS-regression: a basic tool of chemometrics. Chemom Intel Lab Syst 58(2):109–130
Wilson, A. D., & Harwood, L. (2006) Production of monoclonal antibodies specific for native equine IgE and their application to monitor total serum IgE responses in Icelandic and non-Icelandic horses with insect bite dermal hypersensitivity. Vet Immunol Immunopathol, 112: 156–170
Acknowledgements
Nuzul Jambari was sponsored by a full Ph.D. grant from the Ministry of Education of Malaysia.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Jambari, N.N., Wang, X., Alcocer, M. (2017). Protein Microarray-Based IgE Immunoassay for Allergy Diagnosis. 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_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6925-8_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6923-4
Online ISBN: 978-1-4939-6925-8
eBook Packages: Springer Protocols