Nanobody-based fluorescence resonance energy transfer immunoassay for noncompetitive and simultaneous detection of ochratoxin a and ochratoxin B
Nanobody-based fluorescence resonance energy transfer immunoassay for noncompetitive and simultaneous detection of ochratoxin a and ochratoxin B
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基于纳米抗体的荧光共振能量转移免疫分析法，用于非竞争性同时检测赭曲霉毒素 a 和赭曲霉毒素 B
DOI：
10.1016/j.envpol.2019.04.135
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发表时间：
2019-08-01
影响因子：
8.9
通讯作者：
Xu, Yang
中科院分区：
文献类型：
--
作者：
Tang, Zongwen;Liu, Xing;Xu, Yang
利用纳米抗体（Nb）开发了一种非竞争性均相荧光共振能量转移（FRET）免疫分析法，用于高灵敏度地同时检测赭曲霉毒素A（OTA）和赭曲霉毒素B（OTB）。Nb中色氨酸残基（供体）增强的固有荧光（激发波长：280 nm）能够激发OTA和OTB（受体）的荧光用于检测（发射波长：430 nm）。在最佳条件下，基于Nb的FRET免疫分析法对OTA和OTB的检测限分别为0.06 ng/mL和0.12 ng/mL。对OTA和OTB的几种类似物以及非特异性蛋白质和抗体检测到的交叉反应极小。在加标回收研究中获得了可接受的准确度和精密度，并且结果与高效液相色谱法（HPLC）的结果相关性良好。这些结果表明，所开发的方法可作为一种有用的工具，用于非竞争性、均相且同时检测OTA和OTB以及其他具有类似荧光特性的环境分析物。（C）2019爱思唯尔有限公司。保留所有权利。
A noncompetitive and homogeneous fluorescence resonance energy transfer (FRET) immunoassay was developed using a nanobody (Nb) for highly sensitive and simultaneous detection of ochratoxin A (OTA) and ochratoxin B (OTB). The promoted intrinsic fluorescence (lambda(ex): 280 nm) of tryptophan residues (donor) in Nb can excite the fluorescence of OTA and OTB (acceptor) for detection (lambda(em): 430 nm). Using optimal conditions, the limits of detection of the Nb-based FRET immunoassay were 0.06 and 0.12 ng/mL for OTA and OTB, respectively. Minimal cross reactivity was detected for several analogues of OTA and OTB as well as nonspecific proteins and antibodies. Acceptable accuracy and precision were obtained in the spike and recovery study, and the results correlated well with those by HPLC. These results demonstrated that the developed method could be a useful tool for noncompetitive, homogeneous, and simultaneous detection of OTA and OTB as well as other environmental analytes with similar fluorescence properties. (C) 2019 Elsevier Ltd. All rights reserved.