Application of photonic crystal enhanced fluorescence to detection of low serum concentrations of human IgE antibodies specific for a purified cat allergen (Fel D1).
Application of photonic crystal enhanced fluorescence to detection of low serum concentrations of human IgE antibodies specific for a purified cat allergen (Fel D1).
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DOI：
10.1016/j.bios.2015.08.071
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发表时间：
2016-03-15
影响因子：
12.6
通讯作者：
Cunningham BT
中科院分区：
文献类型：
--
作者：
Tan Y;Halsey JF;Tang T;Wetering SV;Taine E;Cleve MV;Cunningham BT
我们证明了使用光子晶体增强荧光（PCEF）微阵列平台检测低浓度的过敏原特异性免疫球蛋白E（IgE）在人血清中。光子晶体（PC）表面被设计为提供Cy 5的激发波长和发射波长的光学共振，用于放大从多路复用过敏原微阵列测量的荧光信号强度。基于表面的夹心免疫测定用于检测和定量针对高度纯化的猫过敏原（Fel D1）的特异性IgE抗体。通过PC微阵列系统和市售临床分析仪测量的IgE的最低可检测浓度的比较表明，PCEF微阵列系统提供更高的灵敏度。PCEF系统能够检测低浓度的特异性IgE（约0.02 kU/L），其灵敏度是市售FDA批准的分析仪的5至17倍。在使用多过敏原阵列的初步实验中，我们证明了选择性同时检测多种过敏原的IgE抗体。
We demonstrate the detection of low concentrations of allergen-specific Immunoglobulin E (IgE) in human sera using a Photonic Crystal Enhanced Fluorescence (PCEF) microarray platform. The Photonic Crystal (PC) surface, designed to provide optical resonances for the excitation wavelength and emission wavelength of Cy5, was used to amplify the fluorescence signal intensity measured from a multiplexed allergen microarray. Surface-based sandwich immunoassays were used to detect and quantify specific IgE antibodies against a highly purified cat allergen (Fel d1). A comparison of the lowest detectable concentration of IgE measured by the PC microarray system and a commercially available clinical analyzer demonstrated that the PCEF microarray system provides higher sensitivity. The PCEF system was able to detect low concentrations of specific IgE (~0.02 kU/L), which is 5 to 17 -fold more sensitive than the commercially available FDA-approved analyzers. In preliminary experiments using multi-allergen arrays, we demonstrate selective simultaneous detection of IgE antibodies to multiple allergens.