Fluorometric flow-immunoassay for alkylphenol polyethoxylates on a microchip containing a fluorescence detector comprised of an organic light emitting diode and an organic photodiode
Fluorometric flow-immunoassay for alkylphenol polyethoxylates on a microchip containing a fluorescence detector comprised of an organic light emitting diode and an organic photodiode
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DOI：
10.1016/j.talanta.2014.10.055
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发表时间：
2015-03-01
期刊：
影响因子：
6.1
通讯作者：
Imato, Toshihiko
中科院分区：
文献类型：
--
作者：
Liu, Rong;Ishimatsu, Ryoichi;Imato, Toshihiko
以有机发光二极管（OLED）为光源，有机光电二极管（OPD）为检测器，在微芯片上构建了一个小型荧光检测器，并用于烷基酚聚氧乙烯醚（APE）的免疫分析。基于铽配合物的OLED在546 nm的主波长处发射尖锐的光，半峰全宽为9 nm。用富勒烯70（C-70）和三[4-（5-苯基硫杂环戊-2-基）苯胺]（TPTPA）制备的OPD在586 nm波长下的入射光电转换效率（IPCE）约为44%。评价荧光检测器测定试卤灵（λ（em）= 586 nm）的性能，由于试卤灵荧光引起的OPD光电流与试卤灵浓度在0 - 18 μ M范围内成正比，检测限（S/N=3）为0.6 μ M。将抗APE抗体固定在聚二甲基硅氧烷（PDMS）微芯片的通道表面或磁性微珠表面，成功地将荧光检测器用于APE的竞争性酶联免疫吸附测定。在与含有辣根过氧化物酶（HRP）标记的APE的APE样品溶液进行免疫反应后，使用荧光检测器测量在引入Amplex Red和H2 O2的混合溶液后立即产生的试卤灵的荧光。由于试卤灵的荧光引起的OPD的光电流信号相对于APE的对数浓度的校准曲线是S形的。定义为IC 80的检测限为约100。1 ppb和ca。对于使用固定在微通道表面上的抗APE抗体的方法和在抗体固定在磁性微珠表面上的情况，分别为2 ppb。(C)2014爱思唯尔有限公司版权所有。
A compact fluorescence detector was constructed on a microchip from an organic light emitting diode (OLED) as the light source and an organic photodiode (OPD) as the photo-detector and was used in an immunoassay for allcylphenol polyethoxylates (APE). The OLED based on a terbium complex emitted a sharp light at the main wavelength of 546 nm with a full width at half maximum of 9 nm. The incident photo-to-current conversion efficiency (IPCE) of the OPD fabricated with Fullerene 70(C-70) and tris[4-(5phenylthiopen-2-yephenyll-amine (TPTPA) was approximately 44% for light at a wavelength of 586 nm. The performance of the fluorescence detector was evaluated for the determination of resorufin (lambda(em)= 586 nm) and the photocurrent of the OPD due to the fluorescence of resorufin was proportional to the concentration of resorufin in the range from 0 to 18 mu M with a detection limit (S/N=3) of 0.6 mu M. The fluorescence detector was successfully utilized in a competitive enzyme-linked immunosorbent assay for APE, where an anti-APE antibody was immobilized on the surface of the channel of the Polydimethylsiloxane (PDMS) microchip or on the surface of magnetic microbeads. After an immunoreaction with a sample solution of APE containing a horse radish peroxidase (HRP)-labeled APE, the fluorescence of resorufin generated just after introduction of a mixed solution of Amplex Red and H2O2 was measured using the fluorescence detector. The calibration curve for the photocurrent signals of the OPD due to the fluorescence of resorufin against the logarithmic concentration of APE was sigmoidal in shape. The detection limits defined as IC80 were ca. 1 ppb and ca. 2 ppb, respectively, for the methods using the anti-APE antibody immobilized on the surface of the microchannel and in the case where the antibody was immobilized on the surface of magnetic microbeads. (C) 2014 Elsevier B.V. All rights reserved.