Environmental and occupational respiratory disordersSimultaneous detection of total and allergen-specific IgE by using purified allergens in a fluorescent multiplex array
Section snippets
Human sera
Human sera originated from studies of allergic sensitization performed previously at the University of Virginia15, 16, 17 (n = 62) or were kindly provided by Dr Ronald van Ree from the European Union Development of certified reference materials for allergenic products and validation of methods for their quantification Allergen Standardization Project (n = 33)18 and by Dr Enrique Fernandez-Caldas of Laboratorios LETI, Spain19 (n = 63). Collection of sera was approved by the institutional review
Array for total IgE
To establish the best coupling concentration of IgE capture mAb in the array for detection of total IgE, activated microspheres were covalently coupled with capture mAb at concentrations of 50 to 800 μg per coupling batch (1.25 × 106 beads). A 2B12-IgE standard curve ranging from 0.05 to 1000 IU IgE/mL was prepared on each of the coupled bead sets. A coupling concentration of 400 μg was found optimal on the basis of signal intensity and dynamic range of the standard curve (Fig 1, A). After
Discussion
The study presented here used fluorescent multiplex suspension array technology to develop a method enabling the simultaneous detection of total and allergen-specific IgE. The benefits of multiplex technology over ELISA methods for various analytes have recently been reviewed.5, 25, 26 Speed, the ability to measure different antibody concentrations with minimum sample volume, and more desirable reaction kinetics and enhanced dynamic range are major benefits of multiplex technology versus ELISA.
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Cited by (53)
Evolution Toward Chip-Based Arrays in the Laboratory Diagnosis of Human Allergic Disease
2023, Journal of Allergy and Clinical Immunology: In PracticeThe use of microarray and other multiplex technologies in the diagnosis of allergy
2021, Annals of Allergy, Asthma and ImmunologyArray-based measurements of aero-allergen-specific IgE correlate with skin-prick test reactivity in asthma regardless of specific IgG4 or total IgE measurements
2021, Journal of Immunological MethodsCitation Excerpt :IgE measurement requires particularly accurate and sensitive assays as its serum concentration is typically low. Established assays for the detection and quantification of total and specific IgE include the radio-allergosorbent test (RAST) and ELISAs but, more recently, protein microarray technology has been applied to the detection and quantification of total and allergen-specific IgE (Bacarese-Hamilton et al., 2002; Fall et al., 2003; Jahn-Schmid et al., 2003; Deinhofer et al., 2004; King et al., 2007) with continued developments both commercially and academically (Melioli et al., 2011; Lupinek et al., 2014; Skrindo et al., 2015). We previously reported the development of a autoantigen-specific microarray for the detection of autoantibodies (Shindi et al., 2017) and report here the development and application of a serological allergen microarray using commercial skin-prick test allergen preparations as the target antigens: these are all purified, natural allergens that detect allergen-specific IgE (sIgE) rather than recombinant antigens that may have structural and antigenic alterations.
A new optical interferometric-based in vitro detection system for the specific IgE detection in serum of the main peach allergen
2020, Biosensors and BioelectronicsCitation Excerpt :Therefore, despite the worldwide acceptance of the CAP as the gold standard for this type of test (Johansson, 2004), there is an urgent need to develop reliable and comparable multiple testing assays to allow the reduction of sample volume, testing time, and costs (Hamilton RG, 2010; Maloney et al., 2008). In this context, new IgE assays such as microarrays and microbeads are addressing clinician requirements for multiplex testing with low volume of sample (Chinnasamy et al., 2014; Gadisseur et al., 2011; Gantelius et al., 2011; King et al., 2007; Lupinek et al., 2014; Pomponi et al., 2012; Ramachandran et al., 2013; van Hage et al., 2017b). In addition, new approaches are combining micro and nanotechnology to manufacture and characterize nanomaterials in order to enhance the sensitivity and accuracy of IgE biosensors (Erathodiyil and Ying, 2011; Jain et al., 2007).
Biosensors for food allergy detection according to specific IgE levels in serum
2020, TrAC - Trends in Analytical ChemistryCitation Excerpt :After immobilisation, the sample is incubated on the chip and the spot intensities produced by fluorophores coupled to the secondary antibody are recorded. Several examples have been published [31–33]. A colorimetric microarray assay based on using a substrate of alkaline phosphatase as the developer reagent has also been developed [34].
Supported in part by Small Business Innovation Research contract ES55545 from the National Institute of Environmental Health Sciences.
Disclosure of potential conflict of interest: E.-M. King, L. D. Vailes, and A. Tsay have received grant support from a National Institute of Environmental Health Sciences Small Business Innovation Research contract and are employed by Indoor Biotechnologies Inc. M. D. Chapman owns stock in and is employed by Indoor Biotechnologies Inc and has received grant support from the National Institute of Environmental Health Sciences. S. M. Santiover has declared that she has no conflict of interest.