Optimization of Critical Factors Affecting the Performance of an Allergen Chip for the Analysis of an Allergen-specific Human IgE in Serum

A sensitive and multiplexed assay of allergen-specific human immunoglobulin E (IgE) is of great significance in the precise diagnosis of allergies. We report on the optimization of critical factors for chip-based analysis of IgE in human serum with a high reliability. Extracts of two mite species we...

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Bibliographic Details
Published in:Analytical Sciences 2007, Vol.23(5), pp.545-549
Main Authors: HAN, Min-Kyu, SEO, Moon-Hyeong, LEE, Dohoon, KIM, Seung Hyun, PARK, Hae-Sim, KIM, Hak-Sung
Format: Article
Language:English
Subjects:
Allergens - immunology
Animals
Antibody Specificity
Fluorescent Antibody Technique - methods
Humans
Immunoglobulin E - blood
Immunoglobulin E - immunology
Protein Array Analysis - methods
Pyroglyphidae - immunology
Sensitivity and Specificity
Species Specificity
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Summary:A sensitive and multiplexed assay of allergen-specific human immunoglobulin E (IgE) is of great significance in the precise diagnosis of allergies. We report on the optimization of critical factors for chip-based analysis of IgE in human serum with a high reliability. Extracts of two mite species were used as model allergens, and were spotted onto a glass slide for the construction of an allergen chip. Respective allergen-specific IgE in human serum was analyzed by using biotinylated anti-human IgE and a streptavidin-Cy3 conjugate. Factors affecting the performance of the allergen chip were investigated and optimized. Especially, the effect of additives, the concentrations of biotinylated anti-human IgE and the streptavidin-Cy3 conjugate, the serum dilution factor, and the concentration of allergen extract as a capturing agent were examined in detail. Under the optimized conditions, a chip-based analysis for sera from 43 patients revealed a reliable and reproducible diagnosis of respective allergies, showing a good correlation with a conventional CAP assay.
ISSN:0910-6340
1348-2246
DOI:10.2116/analsci.23.545