Preparation of Antibodies to Guinea Pig IgE and Its Use for Enzyme-Linked Immunosorbent Assay of IgE Antibodies

Itō, Kōji; Yamauchi, Norihiro; Shoji, Shunsuke; Miyamoto, Yasubumi; Miyamoto, Tomofumi; Watanabe, Naohiro

doi:10.1159/000233823

Cited by 8 publications

(14 citation statements)

References 2 publications

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“…Antigen binding activity of both antibodies in the serum will give us such information. Recently we could obtain rabbit anti-guinea pig IgE [12]. This antiserum may enable these studies to be carried out.…”

Section: Discussionmentioning

confidence: 99%

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IgG2 Antibodies Block IgE Antibody-Induced Asthma in Guinea Pigs

Yamauchi¹,

Itō²,

Suko³

et al. 1986

Int Arch Allergy Immunol

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In order to examine the blocking activity of IgG2 antibodies to guinea pig for IgE antibodies-induced guinea pig asthma, experiments were carried out as follows. Guinea pigs were passively sensitized intravenously with guinea pig serum containing IgE antibodies to ovalbumin (OA). 8 days after sensitization, IgG2 purified from guinea pigs hyperimmunized with OA was intravenously injected. One hour later, the guinea pigs were challenged by inhalation of OA solution. Asthma attacks were not observed in the guinea pigs, whereas the attacks were observed in guinea pigs passively sensitized with the IgE antibodies but injected IgG2 fraction from normal guinea pigs 1 h before inhalation. These observations suggested that IgG antibodies that increased after immunotherapy might block asthma caused by inhalation of allergens in humans.

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Section: Discussionmentioning

confidence: 99%

“…The other 5 sensitized guinea pigs showed a similar reaction at the inhalation of OA. was observed when examined by radioimmuoelectrophoresis [12].…”

Section: Characteristic O F Gp-oa Ige Preparedmentioning

confidence: 99%

IgG2 Antibodies Block IgE Antibody-Induced Asthma in Guinea Pigs

Yamauchi¹,

Itō²,

Suko³

et al. 1986

Int Arch Allergy Immunol

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“…The ELISA was also validated using serum samples from normal and OVA-sensitized guinea pigs for measurement of the IgE mass concentrations. In a 1985 report based on preliminary experiments, Ito et al showed that a rabbit anti-guinea-pig IgE polyclonal antibody could be raised using IgE-rich fraction purified from the pooled serum of guinea pigs infected with Trichinella spiralis as an immunogen and also demonstrated the possibility of establishing an ELISA for guinea pig IgE (22). They also reported development of an ELISA using OVA/ mite extract-coated wells to measure the levels of IgE antibodies to OVA / mite for comparison with titers determined by PCA (11).…”

Section: Discussionmentioning

confidence: 99%

“…Guinea-pig IgE was purified from pooled sera of guinea pigs (female Hartley, about 600 g of body weight; SLC Japan, Inc., Hamamatsu) that had been sensitized eight times at two-week intervals with 10 µg of ovalbumin (OVA; ICN Biochemicals, Aurora, OH, USA) in 4 mL of aluminum hydroxide gel prepared in our laboratory and 2.5 × 10 10 cells of Pertussis Vaccine (Wako Pure Chemical Industries Ltd., Osaka) per animal based on Ito's method with some modifications (11,22). The method for purification consisted of salting-out with saturated ammonium sulfate and ion-exchange chromatography.…”

Section: Standard Guinea Pig-igementioning

confidence: 99%

“…On the other hand, there is no practical method available for a multisample quantitative assay of guinea-pig IgE except for PCA, although some reports have been published on the determination of guinea-pig IgE without using a standard material (11,22). It has therefore been hoped that there would be development of a practical immunoassay with high sensitivity and specificity for quantification of the guinea-pig IgE mass concentration.…”

Section: Introductionmentioning

confidence: 99%

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Development and Evaluation of a Direct Sandwich Enzyme-Linked Immunosorbent Assay for the Quantification of Guinea-Pig Immunoglobulin E

et al. 2006

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Abstract. The purpose of this study was to develop and evaluate a direct sandwich enzymelinked immunosorbent assay (ELISA) for the immunoglobulin E (IgE) in serum and plasma from guinea pig using mouse monoclonal antibodies specific for guinea-pig IgE. Mouse monoclonal antibodies were raised against purified IgE protein. The ELISA was performed using a combination of two anti-IgE monoclonal antibodies. One antibody was labeled with horseradish peroxidase (HRP), and the other was coated on polystyrene wells. Purified guinea-pig IgE was used as the standard material. The validity of the ELISA was confirmed by precision, dilution, recovery, and interference tests. The range of detection was 3.1 -800 ng of IgE mass per mL of serum and plasma. The intra-and inter-assay coefficients of variation were 4.6% and 5.7%, respectively, or less. The recovery test showed variation only between 92.1% and 111.8%, and the anticoagulants showed noninterference with the IgE assay. The mean serum IgE mass concentration in OVA-sensitized guinea pigs was 29438 ng / mL, and it was 48.6 ng / mL in normal guinea pigs. The present ELISA is useful and practical for specific measurement of the guinea-pig IgE, and it is surmised that it would be suitable for use in allergological and pharmacological research.

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