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

Young Ki Hahn, Zongwen Jin, Joo H. Kang, Eunkeu Oh, Min Kyu Han, Hak Sung Kim, Jung Tak Jang, Jae-Hyun Lee, Jinwoo Cheon, Seung Hyun Kim, Hae Sim Park, Je Kyun Park

Research output: Contribution to journalArticle

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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 (∼104 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 (∼10 μL).

Original languageEnglish
Pages (from-to)2214-2220
Number of pages7
JournalAnalytical Chemistry
Volume79
Issue number6
DOIs
Publication statusPublished - 2007 Mar 15

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Allergens
Immunoglobulin E
Magnetic fields
Microchannels
Assays
Nanoparticles
Polystyrenes
Microfluidics
Permanent magnets
Dust
Microstructure

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Hahn, Y. K., Jin, Z., Kang, J. H., Oh, E., Han, M. K., Kim, H. S., ... Park, J. K. (2007). Magnetophoretic immunoassay of allergen-specific IgE in an enhanced magnetic field gradient. Analytical Chemistry, 79(6), 2214-2220. https://doi.org/10.1021/ac061522l
Hahn, Young Ki ; Jin, Zongwen ; Kang, Joo H. ; Oh, Eunkeu ; Han, Min Kyu ; Kim, Hak Sung ; Jang, Jung Tak ; Lee, Jae-Hyun ; Cheon, Jinwoo ; Kim, Seung Hyun ; Park, Hae Sim ; Park, Je Kyun. / Magnetophoretic immunoassay of allergen-specific IgE in an enhanced magnetic field gradient. In: Analytical Chemistry. 2007 ; Vol. 79, No. 6. pp. 2214-2220.
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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 (∼104 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 (∼10 μL).",
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Hahn, YK, Jin, Z, Kang, JH, Oh, E, Han, MK, Kim, HS, Jang, JT, Lee, J-H, Cheon, J, Kim, SH, Park, HS & Park, JK 2007, 'Magnetophoretic immunoassay of allergen-specific IgE in an enhanced magnetic field gradient', Analytical Chemistry, vol. 79, no. 6, pp. 2214-2220. https://doi.org/10.1021/ac061522l

Magnetophoretic immunoassay of allergen-specific IgE in an enhanced magnetic field gradient. / Hahn, Young Ki; Jin, Zongwen; Kang, Joo H.; Oh, Eunkeu; Han, Min Kyu; Kim, Hak Sung; Jang, Jung Tak; Lee, Jae-Hyun; Cheon, Jinwoo; Kim, Seung Hyun; Park, Hae Sim; Park, Je Kyun.

In: Analytical Chemistry, Vol. 79, No. 6, 15.03.2007, p. 2214-2220.

Research output: Contribution to journalArticle

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T1 - Magnetophoretic immunoassay of allergen-specific IgE in an enhanced magnetic field gradient

AU - Hahn, Young Ki

AU - Jin, Zongwen

AU - Kang, Joo H.

AU - Oh, Eunkeu

AU - Han, Min Kyu

AU - Kim, Hak Sung

AU - Jang, Jung Tak

AU - Lee, Jae-Hyun

AU - Cheon, Jinwoo

AU - Kim, Seung Hyun

AU - Park, Hae Sim

AU - Park, Je Kyun

PY - 2007/3/15

Y1 - 2007/3/15

N2 - 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 (∼104 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 (∼10 μL).

AB - 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 (∼104 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 (∼10 μL).

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DO - 10.1021/ac061522l

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EP - 2220

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

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