Use of carbon quantum dots and fluorescein isothiocyanate in developing an improved competitive fluoroimmunoassay for detecting polybrominated diphenyl ether
Use of carbon quantum dots and fluorescein isothiocyanate in developing an improved competitive fluoroimmunoassay for detecting polybrominated diphenyl ether
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使用碳量子点和异硫氰酸荧光素开发改进的竞争性荧光免疫测定法来检测多溴二苯醚
DOI：
10.1007/s13738-019-01639-w
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发表时间：
2019-08-01
影响因子：
2.4
通讯作者：
Cai，Qingyun
中科院分区：
文献类型：
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作者：
Li，Weili;Yao，Lu;Cai，Qingyun
多溴二苯醚 (PBDE) 在多种消费品中用作阻燃剂。一旦释放到环境中，多溴联苯醚就被称为“持久性有机污染物”，具有生物累积性，导致人类和生态系统在接触后遭受不良健康影响。在这项工作中，提出了两种使用竞争性荧光免疫分析 (FIA) 检测 PBDE 的策略。为了开发 PBDE 的竞争性 FIA，我们实验室选择了模型 PBDE 化合物 2, 3', 4, 5', 6-五溴二苯醚 (BDE-121) 进行抗体开发，并通过半抗原合成制备了针对 BDE-121 的单克隆抗体。采用碳量子点 (CQD) 和异硫氰酸荧光素 (FITC) 作为荧光标记来生产 FIA。实验结果表明，生物缀合物CQDs-BSA-BDE-121和FITC-BSA-BDE-121在检测BDE-121方面均表现出良好的灵敏度和选择性，因此都是有效的策略。然而，还确定 CQD 的性能优于 FITC，这可能是由于其卓越的生物相容性和光稳定性，显示出 0.5 nM–100 nM (0.28 ng/mL ~ 56.47 ng/mL) 的线性范围，检测限为 0.1 nM (0.056 ng/mL)。因此，这项研究证实了将载体蛋白/半抗原与生物修饰的量子点相结合来检测小分析物的潜力。最后，CQDs和FITC被证明是荧光共振能量转移（FRET）的优质供体/受体对，这将为研究基于CQDs和FITC的FRET用于PBDE检测和定量提供途径和基础。
Polybrominated diphenyl ethers (PBDEs) are used as a flame retardant in a wide variety of consumer products. Once released into the environment, PBDEs are characterized as “Persistent Organic Pollutants” that have an affinity for bioaccumulation, resulting in humans and the ecosystem suffering ill health effects upon exposure. In this work, two strategies are proposed to detect PBDEs using competitive fluoroimmunoassay (FIA). To develop a competitive FIA of PBDEs, a model PBDE compound, 2, 3′, 4, 5′, 6-pentabromodiphenylether (BDE-121), was selected for antibody development, and a monoclonal antibody against BDE-121 was prepared, via hapten synthesis, in our laboratory. Carbon quantum dots (CQDs) and fluorescein isothiocyanate (FITC) were employed as fluorescent labels to produce the FIA. Experimental results showed that the bioconjugates CQDs-BSA-BDE-121 and FITC-BSA-BDE-121 both demonstrate good sensitivity and selectivity for detecting BDE-121 and are therefore both effective strategies. However, it was also determined that CQDs performed better than the FITC, likely due to their superior biocompatibility and photostability, displaying a linear range of 0.5 nM–100 nM (0.28 ng/mL ~ 56.47 ng/mL) with a limit of detection of 0.1 nM (0.056 ng/mL). Thus, this study confirmed the potential of combining carrier protein/haptens with biologically modified QDs to detect small analytes. Finally, CQDs and FITC were shown to be a good-quality donor/acceptor pair for Fluorescence Resonance Energy Transfer (FRET), which will provide a path and foundation for researching FRET based on CQDs and FITC for PBDE detection and quantification.