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Ultrafast Multiplexed-Allergen Detection through Advanced Fluidic Design and Monolithic Interferometric Silicon Chips
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-07-12 00:00:00 , DOI: 10.1021/acs.analchem.8b02321 Michailia Angelopoulou 1 , Panagiota S. Petrou 1 , Eleni Makarona 2 , Willem Haasnoot 3 , Isabella Moser 4 , Gerhard Jobst 4 , Dimitrios Goustouridis 5, 6 , Michèle Lees 7 , Kalliopi Kalatzi 8 , Ioannis Raptis 2 , Konstantinos Misiakos 2 , Sotirios E. Kakabakos 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-07-12 00:00:00 , DOI: 10.1021/acs.analchem.8b02321 Michailia Angelopoulou 1 , Panagiota S. Petrou 1 , Eleni Makarona 2 , Willem Haasnoot 3 , Isabella Moser 4 , Gerhard Jobst 4 , Dimitrios Goustouridis 5, 6 , Michèle Lees 7 , Kalliopi Kalatzi 8 , Ioannis Raptis 2 , Konstantinos Misiakos 2 , Sotirios E. Kakabakos 1
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A silicon-based miniaturized sensor chip combined with an advanced microfluidic module for the simultaneous, label-free immunochemical determination of four allergens, bovine milk protein, peanut protein, soy protein, and gliadin, is presented. The sensor chip consists of an array of 10 broad-band Mach–Zehnder interferometers (BB-MZIs) monolithically integrated on silicon, along with their respective broad-band light sources. The BB-MZIs were biofunctionalized with the targeted allergens and their responses during immunoreaction were monitored by multiplexing their transmission spectra through an external miniaturized spectrometer. The assay is performed by running mixtures of calibrators or samples with the antibodies against the four allergens followed by an antispecies specific antibodies solution. Employing a fluidic module of nearly one-dimensional geometry, that provided for uniform delivery of the reagents, CV values <6% were achieved for the responses of the 10 BB-MZIs, allowing for reliable multianalyte determinations. The analysis is completed in 6.5 min, and the detection limits were 0.04 μg/mL for bovine k-casein, 1.0 μg/mL for peanut protein, 0.80 μg/mL for soy protein, and 0.10 μg/mL for gliadin. The assays were accurate (recoveries 88–118%) and repeatable (intra- and interassay CVs <7% for all four allergens). Finally, the sensor was evaluated by analyzing samples from a cleaning in place system (CIP) of a dairy industry and the results obtained were in good agreement with those received by the respective ELISAs. The analytical characteristics of the sensor combined with the short analysis time and the small chip size make the proposed system an ideal tool for on-site multianalyte determinations.
中文翻译：
通过先进的流体设计和单片干涉硅芯片实现超快速多路复用变应原检测
提出了一种基于硅的微型传感器芯片，结合了先进的微流控模块，可同时，无标记地对四种过敏原（牛乳蛋白，花生蛋白，大豆蛋白和麦醇溶蛋白）进行免疫化学测定。传感器芯片由单片集成在硅片上的10个宽带马赫曾德尔干涉仪（BB-MZIs）阵列以及它们各自的宽带光源组成。BB-MZIs通过目标过敏原进行了生物功能化，并通过外部微型光谱仪对它们的透射光谱进行了多路复用，从而监测了其在免疫反应过程中的反应。通过运行校准物或样品与针对四种过敏原的抗体的混合物，然后是抗物种特异性抗体溶液，进行分析。采用几乎一维几何形状的流体模块，该模块可实现试剂的均匀输送，对于10个BB-MZI的响应，CV值<6％，可进行可靠的多分析物测定。分析在6.5分钟内完成，牛k-酪蛋白的检出限为0.04μg/ mL，花生蛋白的检出限为1.0μg/ mL，大豆蛋白的检出限为0.80μg/ mL，而麦醇溶蛋白的检出限为0.10μg/ mL。该测定准确（回收率88–118％）和可重复（所有四种过敏原的测定内和测定间CV <7％）。最后，通过分析来自乳制品业的就地清洗系统（CIP）的样品来评估传感器，并且所获得的结果与相应ELISA所获得的结果高度吻合。
更新日期：2018-07-12
中文翻译：
通过先进的流体设计和单片干涉硅芯片实现超快速多路复用变应原检测
提出了一种基于硅的微型传感器芯片，结合了先进的微流控模块，可同时，无标记地对四种过敏原（牛乳蛋白，花生蛋白，大豆蛋白和麦醇溶蛋白）进行免疫化学测定。传感器芯片由单片集成在硅片上的10个宽带马赫曾德尔干涉仪（BB-MZIs）阵列以及它们各自的宽带光源组成。BB-MZIs通过目标过敏原进行了生物功能化，并通过外部微型光谱仪对它们的透射光谱进行了多路复用，从而监测了其在免疫反应过程中的反应。通过运行校准物或样品与针对四种过敏原的抗体的混合物，然后是抗物种特异性抗体溶液，进行分析。采用几乎一维几何形状的流体模块，该模块可实现试剂的均匀输送，对于10个BB-MZI的响应，CV值<6％，可进行可靠的多分析物测定。分析在6.5分钟内完成，牛k-酪蛋白的检出限为0.04μg/ mL，花生蛋白的检出限为1.0μg/ mL，大豆蛋白的检出限为0.80μg/ mL，而麦醇溶蛋白的检出限为0.10μg/ mL。该测定准确（回收率88–118％）和可重复（所有四种过敏原的测定内和测定间CV <7％）。最后，通过分析来自乳制品业的就地清洗系统（CIP）的样品来评估传感器，并且所获得的结果与相应ELISA所获得的结果高度吻合。
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