The IgE-facilitated allergen binding (FAB) assay:: Validation of a novel flow-cytometric based method for the detection of inhibitory antibody responses
The IgE-facilitated allergen binding (FAB) assay:: Validation of a novel flow-cytometric based method for the detection of inhibitory antibody responses
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DOI：
10.1016/j.jim.2006.09.004
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发表时间：
2006-12-20
影响因子：
2.2
通讯作者：
Francis, James N.
中科院分区：
文献类型：
--
作者：
Shamji, Mohamed H.;Wilcock, Louisa K.;Francis, James N.
IgE 促进过敏原结合 (IgE-FAB) 测定代表了促进过敏原呈递的体外模型。将过敏原-IgE 复合物与 EBV 转化的 B 细胞系一起孵育，并通过流式细胞术检测与细胞表面 CD23 结合的复合物。先前已证明，添加接受过过敏原特异性免疫治疗的患者血清可抑制过敏原-IgE 复合物与 B 细胞上 CD23 的结合。在这项研究中，我们根据国际协调会议的指南描述了草花粉特异性 IgE-FAB 测定的表征和分析验证。我们确定了 IgE-FAB 测定内和测定间的变异性，并定义了该测定的检测限。我们还证明了测定的线性和稳健性。利用草花粉免疫治疗的随机双盲安慰剂对照试验（n = 33）的结果，我们使用 ROC 曲线分析定义了 IgE-FAB 测定的临床敏感性和特异性。总之，IgE-FAB 测定具有可重复性、稳健性、敏感性，并且是一种适合作为监测接受过敏原免疫治疗的患者的抑制性抗体功能的工具的特定方法。 (c) 2006 Elsevier B.V. 保留所有权利。
The IgE-facilitated allergen binding (IgE-FAB) assay represents an in vitro model of facilitated allergen presentation. Allergen-IgE complexes are incubated with an EBV-transformed B cell line and complexes bound to CD23 on the surface of cells are detected by flow cytometry. The addition of serum from patients who have received allergen-specific immunotherapy has been shown previously to inhibit allergen-IgE complex binding to CD23 on B cells.In this study, we describe the characterisation and analytical validation of the grass pollen-specific IgE-FAB assay according to guidelines from the International Conference on Harmonisation. We established the intra- and inter-assay variability of IgE-FAB and have defined the detection limits of this assay. We have also demonstrated assay linearity and robustness. Using the results from a randomised double-blind placebo-controlled trial of grass pollen immunotherapy (n=33), we have defined the clinical sensitivity and specificity of the IgE-FAB assay using ROC curve analysis.In conclusion, the IgE-FAB assay is reproducible, robust, sensitive and a specific method suitable as a tool for monitoring inhibitory antibody function from patients receiving allergen immunotherapy. (c) 2006 Elsevier B.V. All rights reserved.