Event Abstract

Multiplex immunoassay for allergens using oligonucleotide tethering


Allergy to cow’s milk affects about 2.5% of all children under age 3. While most infants overcome this allergy, milk intolerance is prevalent in most countries. Milk allergy is often associated with other food allergy, as well as, respiratory allergy.
Microplate-based multiplexed immunoassays are useful for the simultaneous measure of multiple analytes from a single sample in a single well. This allows for the creation of disease-specific panels potentially leading to a more accurate prognosis and/or diagnosis of the disease state. The A-squared (A2) MicroArray System (QuantiScientifics, Irvine, CA, USA) is a microplate-based platform for quantitative multiplexed immunoassays. Arrays of oligonucleotides are covalently attached to the bottom of the plate well. Complementary strands are in turn linked to antibodies or antigens. The prepared oligo-antibody (-antigen) conjugates are pooled and applied to the wells. Through the process of hybridization self-assembly of the hybrids leads to the creation of a diagnostic multiplex panel in each well of the plate. In this study, several milk proteins recognized as allergens were assembled into a multiplex panel using oligonucleotide tethering. Here, purified milk proteins whose epitopes are known to be associated with the allergy, as well as, crude milk allergen extracts are conjugated to oligonucleotides to create the panel. The panel included F2 crude extract, beta-lactoglobulin, alpha-lactalbumin, IgG, bovine serum albumin, lactoferrin, and 3 caseins. 13 patient serums were screened for the presence of specific IgE providing a profile and subsequent identification of principal component allergen.

Keywords: Oligonucleotides, arrays, Multiplex assay, Allergens, immunoassays

Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.

Presentation Type: Abstract

Topic: Immune-mediated disease pathogenesis

Citation: Matson R (2013). Multiplex immunoassay for allergens using oligonucleotide tethering
. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00052

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Received: 09 Mar 2013; Published Online: 22 Aug 2013.

* Correspondence: Dr. Robert Matson, QUANTISCIENTIFICS LLC, Irvine, California, United States, rsmatson@quantiscientifics.com