A displacement flow immunosensor for explosive detection using microcapillaries
A displacement flow immunosensor for explosive detection using microcapillaries
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DOI：
10.1021/ac970153d
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发表时间：
1997-07-15
影响因子：
7.4
通讯作者：
Ligler, FS
中科院分区：
文献类型：
--
作者：
Narang, U;Gauger, PR;Ligler, FS
在本文中，我们报告了一种基于毛细管位移流免疫传感器的爆炸物，2,4,6-三硝基甲苯（TNT）的表征。使用异双功能交联剂将抗tnt抗体固定在熔融二氧化硅毛细管的硅化内壁上，然后用荧光标记的抗原使毛细管饱和。为了进行检测，将等分的目标分析物溶液注入毛细管柱。目标分析物竞争荧光团标记抗原的结合袋，这是下游检测到的。我们已经证明，与填充柱或基于膜的位移免疫传感器相比，基于毛细管的免疫传感器的灵敏度提高了至少3个数量级，我们已经研究了毛细管的流速和长度对毛细管免疫传感器性能的影响。我们已经证明，通过适当选择抗体，可以制备用于TNT特异性检测的毛细管免疫传感器，或者作为TNT及其主要代谢物的检测器。毛细管免疫传感器对TNT的检出限为440 amol （100 μ L注射1 pg/mL TNT溶液），连续10次向毛细管免疫传感器注射5 ng/mL TNT，其峰下面积的相对标准偏差（RSD）为4%。柱间变异的RSD为5% （n = 5）。整个分析可以在不到3分钟内完成。
In this paper, we report on the characterization of a capillary-based displacement flow immunosensor for the explosive, 2,4,6-trinitrotoluene (TNT). Anti-TNT antibody is immobilized onto the silanized inner walls of a fused-silica capillary using a heterobifunctional cross-linker followed by saturating the capillary with fluorophore-labeled antigen, To perform the assay, an aliquot of target analyte solution is injected into the capillary column. The target analyte competes for the binding pocket of the fluorophore-labeled antigen, which is detected downstream. We have demonstrated that the sensitivity of the capillary-based immunosensor is improved by at least 3 orders of magnitude compared to packed-column or membrane-based displacement now immunosensors, We have investigated the effect of now rate and length of the capillary on the performance of the capillary immunosensor. We have demonstrated that, by the appropriate choice of an antibody, one can prepare a capillary immunosensor for the specific detection of TNT or as a detector for both TNT and its major metabolites. The limit of detection for TNT using the capillary immunosensor is 440 amol (100 mu L injection of 1 pg/mL TNT solution), The relative standard deviation (RSD) of the area under the peaks resulting from 10 back-to-back 100 mu L injections of 5 ng/mL TNT to the capillary immunosensor is 4%. The RSD for column-to-column variation is observed to be 5% (n = 5). The entire assay can be performed in less than 3 min.