Surface plasmon resonance immunosensor for IgE analysis using two types of anti-IgE antibodies with different active recognition sites

Yan Li, Jujie Ren, Hizuru Nakajima, Nobuaki Soh, Koji Nakano, Toshihiko Imato

Research output: Contribution to journalArticle

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Abstract

A simple and novel method for the determination of an IgE antibody based on a surface plasmon resonance immunosensor for the diagnosis of an allergy is described. The method involves the use of an anti-IgE(D) antibody and an anti-IgE(H) antibody, which reacts with the Ce2 domain and the Ce3 domain of the IgE antibody. The anti-IgE(D) antibody was immobilized on the gold surface of a sensor chip by physical adsorption. An IgE antibody sample was incubated by adding it to an anti-IgE(H) antibody solution to form an anti-IgE(H) immunocomplex through a reaction of the Ce3 domain of the IgE antibody. The incubated solution was introduced onto the sensor chip and the immunocomplex of the IgE-anti-IgE(H) then reacted with the anti-IgE(D) antibody immobilized on the sensor chip through the Ce2 domain of the IgE antibody part of the IgE-anti-IgE(H) immunocomplex. The detection limit of the present method for the determination of the IgE antibody was about 10 ppb. The affinity constants for the anti-IgE(H) antibody immunocomplex with the IgE antibody in solution and that of the anti-IgE(H) antibody immunocomplex with the IgE antibody immobilized on the sensor chip by a biotin-streptavidin interaction were estimated to be 4.1 × 10 7 M -1 and 5.8 × 10 6 M -1 , respectively. The affinity constant for the immunocomplex of the anti-IgE(H) antibody with the IgE antibody with the anti-IgE(D) immobilized on the sensor chip was estimated to be 4.9 × 10 7 M -1 , 20-times larger than the affinity constant for the IgE antibody immunocomplex with the anti-IgE(D) antibody immobilized on the sensor chip, based on a direct immunoassay method of the IgE antibody under the same experimental conditions. 2007

Original languageEnglish
Pages (from-to)31-38
Number of pages8
Journalanalytical sciences
Volume23
Issue number1
DOIs
Publication statusPublished - Jan 2007

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Immunosensors
Surface plasmon resonance
Immunoglobulin E
Antibodies
Immobilized Antibodies
Sensors
anti-IgE antibodies
Allergies
Streptavidin
Biotin

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Surface plasmon resonance immunosensor for IgE analysis using two types of anti-IgE antibodies with different active recognition sites. / Li, Yan; Ren, Jujie; Nakajima, Hizuru; Soh, Nobuaki; Nakano, Koji; Imato, Toshihiko.

In: analytical sciences, Vol. 23, No. 1, 01.2007, p. 31-38.

Research output: Contribution to journalArticle

Li, Yan ; Ren, Jujie ; Nakajima, Hizuru ; Soh, Nobuaki ; Nakano, Koji ; Imato, Toshihiko. / Surface plasmon resonance immunosensor for IgE analysis using two types of anti-IgE antibodies with different active recognition sites. In: analytical sciences. 2007 ; Vol. 23, No. 1. pp. 31-38.
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abstract = "A simple and novel method for the determination of an IgE antibody based on a surface plasmon resonance immunosensor for the diagnosis of an allergy is described. The method involves the use of an anti-IgE(D) antibody and an anti-IgE(H) antibody, which reacts with the Ce2 domain and the Ce3 domain of the IgE antibody. The anti-IgE(D) antibody was immobilized on the gold surface of a sensor chip by physical adsorption. An IgE antibody sample was incubated by adding it to an anti-IgE(H) antibody solution to form an anti-IgE(H) immunocomplex through a reaction of the Ce3 domain of the IgE antibody. The incubated solution was introduced onto the sensor chip and the immunocomplex of the IgE-anti-IgE(H) then reacted with the anti-IgE(D) antibody immobilized on the sensor chip through the Ce2 domain of the IgE antibody part of the IgE-anti-IgE(H) immunocomplex. The detection limit of the present method for the determination of the IgE antibody was about 10 ppb. The affinity constants for the anti-IgE(H) antibody immunocomplex with the IgE antibody in solution and that of the anti-IgE(H) antibody immunocomplex with the IgE antibody immobilized on the sensor chip by a biotin-streptavidin interaction were estimated to be 4.1 × 10 7 M -1 and 5.8 × 10 6 M -1 , respectively. The affinity constant for the immunocomplex of the anti-IgE(H) antibody with the IgE antibody with the anti-IgE(D) immobilized on the sensor chip was estimated to be 4.9 × 10 7 M -1 , 20-times larger than the affinity constant for the IgE antibody immunocomplex with the anti-IgE(D) antibody immobilized on the sensor chip, based on a direct immunoassay method of the IgE antibody under the same experimental conditions. 2007",
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