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Magnetophoretic immunoassay of allergen-specific IgE in an enhanced magnetic field gradient.

Authors
Hahn, YK; Jin, Z; Kang, JH; Oh, E; Han, MK; Kim, HS; Jang, JT; Lee, JH; Cheon, J; Kim, SH; Park, HS; Park, JK
Citation
Analytical chemistry, 79(6):2214-2220, 2007
Journal Title
Analytical chemistry
ISSN
0003-27001520-6882
Abstract
We demonstrate a novel magnetophoretic immunoassay of allergen-specific immunoglobulin E (IgE) based on the magnetophoretic deflection velocity of a microbead that is proportional to the associated magnetic nanoparticles under enhanced magnetic field gradient in a microchannel. In this detection scheme, two types of house dust mites, Dermatophagoides farinae (D. farinae) and Dermatophagoides pteronyssinus (D. pteronyssinus), were used as the model allergens. Polystyrene microbeads were conjugated with each of the mite extracts followed by incubation with serum samples. The resulting mixture was then reacted with magnetic nanoparticle-conjugated anti-human IgE for detection of allergen-specific IgE by using sandwich immuno-reactions. A ferromagnetic microstructure combined with a permanent magnet was employed to increase the magnetic field gradient ( approximately 10(4) T/m) in a microfluidic device. The magnetophoretic velocities of microbeads were measured in a microchannel under applied magnetic field, and the averaged velocity was well correlated with the concentration of allergen-specific IgE in serum. From the analysis of pooled sera obtained from 44 patients, the detection limits of the allergen-specific human IgEs for D. farinae and D. pteronyssinus were determined to be 565 (0.045 IU/mL) and 268 fM (0.021 IU/mL), respectively. These values are 1 order of magnitude lower than those by a conventional CAP system. For evaluation of reproducibility and accuracy, unknown sera were subjected to a blind test by using the developed assay system, and they were compared with the CAP system. As a result, coefficient of variance was less than 10%, and the developed method enabled a fast assay with a tiny amount of serum ( approximately 10 microL).
MeSH terms
AdolescentAdultAllergens/immunology*Aminoglycosides/chemistryAnimalsAntibody SpecificityChildChild, PreschoolGoatsHumansImmunoassay/methods*Immunoglobulin E/blood*Immunoglobulin E/immunology*Magnetics*Middle AgedMites/immunology
DOI
10.1021/ac061522l
PMID
17288405
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Allergy
Journal Papers > Research Organization > Regional Clinical Trial Center
AJOU Authors
김, 승현박, 해심
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