Quantitative measurement of IgE antibodies to purified allergens using streptavidin linked to a high-capacity solid phase
Quantitative measurement of IgE antibodies to purified allergens using streptavidin linked to a high-capacity solid phase
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DOI：
10.1016/j.jaci.2004.12.1131
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发表时间：
2005-05-01
影响因子：
14.2
通讯作者：
Platts-Mills, TAE
中科院分区：
文献类型：
--
作者：
Erwin, EA;Custis, NJ;Platts-Mills, TAE
背景：市售的IgE抗体检测方法为许多过敏原提取物提供了每毫升国际单位的结果，但这对于纯化的或新的过敏原来说不容易实现。目的：利用市售的免疫吸附剂，建立适用于纯化过敏原或新过敏原的IgE抗体检测方法。方法：用链亲和素偶联高容量免疫吸附剂（CAP）结合生物素化纯化的螨（Der p1和Der p2）、猫（Fel d1）和狗（Can f1）变应原。对儿童（哮喘和对照组）以及成人特应性皮炎患者的血清进行了这些过敏原的IgE抗体检测。结果：通过连续稀释高、低水平IgE抗体的血清，验证了结果，并采用标准曲线以国际单位/毫升进行定量。对Der p1、Der p2和Fel d1的IgE抗体值与过敏原提取物的值相关(r(2)分别为0.80、0.84和0.95；P < 0.001)。此外，高暴露于螨和猫过敏原的儿童血清中的IgE抗体值表明，Der p1和Der p2的IgE抗体水平比Fel d1高10倍（p < 0.001）。结论：链霉亲和素免疫吸附技术为纯化蛋白的IgE抗体定量提供了一种新的方法。结果为特应性皮炎患者对纯化吸入性过敏原的高量IgE抗体提供了证据。此外，研究结果表明，在高度暴露于螨虫和猫过敏原的儿童中，针对这两种过敏原的IgE抗体存在重大差异。
Background: Commercially available assays for IgE antibody provide results in international units per milliliter for many allergen extracts, but this is not easily achieved with purified or novel allergens. Objective: To develop assays for IgE antibody suitable for purified or novel allergens by using a commercially available immunosorbent.Methods: Streptavidin coupled to a high-capacity immunosorbent (CAP) was used to bind biotinylated purified allergens from mite (Der p 1 and Der p 2), cat (Fel d 1), and dog (Can f 1). Assays for IgE antibody to these allergens were performed on sera from children (asthma and control) as well as adults with atopic dermatitis.Results: The results were validated by serial dilution of sera with high and low levels of IgE antibody and were quantitated in international units per milliliter by using a standard curve. Values for IgE antibody to Der p 1, Der p 2, and Fel d 1 correlated with values obtained with the allergen extracts (r(2) = 0.80, 0.84, and 0.95, respectively; P < .001 in each case). Furthermore, the values for IgE antibody in sera from children with high exposure to mite and cat allergens demonstrated 10-fold higher levels of IgE antibody to Der p 1 and Der p 2 than to Fel d 1 (P < .001).Conclusion: The streptavidin immunosorbent technique provides a new method for quantifying IgE antibody to purified proteins. The results provide evidence about the high quantities of IgE antibody to purified inhalant allergens in patients with atopie dermatitis. In addition, the results demonstrate major differences in IgE antibodies specific for mite and cat allergens among children with high exposure to both allergens.