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Optimization of critical factors affecting the performance of an allergen chip for the analysis of an allergen-specific human IgE in serum.

Authors
Han, MK; Seo, MH; Lee, D; Kim, SH; Park, HS; Kim, HS
Citation
Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 23(5):545-549, 2007
Journal Title
Analytical sciences : the international journal of the Japan Society for Analytical Chemistry
ISSN
0910-63401348-2246
Abstract
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.
MeSH terms
Allergens/immunology*AnimalsAntibody Specificity*Fluorescent Antibody Technique/methods*HumansImmunoglobulin E/blood*Immunoglobulin E/immunology*Protein Array Analysis/methods*Pyroglyphidae/immunologySensitivity and SpecificitySpecies Specificity
PMID
17495399
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Allergy
Journal Papers > Research Organization > Regional Clinical Trial Center
AJOU Authors
김승현박해심
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