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[一种使用单克隆抗体定量检测日本柳杉(Cryptomeria japonica)花粉主要过敏原Cry j I的酶联免疫吸附测定法]

[An enzyme-linked immunosorbent assay for the quantitation of the major allergen from Japanese cedar (Cryptomeria japonica) pollen, Cry j I, using monoclonal antibodies].

作者信息

Watanabe M, Tamura M, Nagoya T, Takahashi Y, Katagiri S, Oka T

机构信息

Kowa Research Institute, Kowa Co., Ltd.

出版信息

Arerugi. 1992 Nov;41(11):1535-9.

PMID:1492786
Abstract

We have developed an enzyme-linked immunosorbent assay (ELISA) using two anti-Cry j I monoclonal antibodies (KW-S10 and KW-S131) for the quantitation of the major allergen from Japanese cedar (Cryptomeria Japonica) pollen, Cry j I. Polystyrene microplates coated with KW-S131 were incubated with pollen allergen extracts. Cry j I, which bound to the antibody, was detected with alkaline phosphatase-conjugated KW-S10 using chromogenic enzyme substrate. Cry j I could be measured in concentration of between 0.16 and 2.5 ng/ml by this assay. Intra- and inter-assay coefficients of variation were 1.1-3.5% and 0.9-4.6%, respectively. This assay was considered that it was specific for Cry j I of Japanese cedar pollen because it didn't react with allergens of hinoki (Chamaecypairs obtusa) pollen which have antigenicity in common to Japanese cedar pollen. This assay would be useful for the standardization of Japanese cedar pollen allergen extracts.

摘要

我们开发了一种酶联免疫吸附测定法(ELISA),使用两种抗Cry j I单克隆抗体(KW-S10和KW-S131)来定量日本柳杉(Cryptomeria Japonica)花粉中的主要过敏原Cry j I。将包被有KW-S131的聚苯乙烯微孔板与花粉过敏原提取物一起孵育。与抗体结合的Cry j I,使用发色酶底物,通过碱性磷酸酶偶联的KW-S10进行检测。通过该测定法,Cry j I的浓度可在0.16至2.5 ng/ml之间进行测量。批内和批间变异系数分别为1.1 - 3.5%和0.9 - 4.6%。该测定法被认为对日本柳杉花粉的Cry j I具有特异性,因为它不与日本扁柏(Chamaecypairs obtusa)花粉的过敏原发生反应,而日本扁柏花粉与日本柳杉花粉具有共同的抗原性。该测定法将有助于日本柳杉花粉过敏原提取物的标准化。

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