Li Taihua, Choi Yo Han, Shin Yong-Beom, Kim Hwa-Jung, Kim Min-Gon
College of Biology and the Environment, Nanjing Forestry University, Nanjing 210-037, China.
Department of Chemistry, School of Physics and Chemistry, Gwangju Institute of Science & Technology (GIST), 261 Cheomdan-gwagiro, Gwangju 500-712, South Korea; Department of Microbiology, College of Medicine, Chungnam National University, Daejeon 301-747, South Korea.
Chemosphere. 2016 May;150:407-413. doi: 10.1016/j.chemosphere.2016.01.008. Epub 2016 Jan 12.
A fluorescence enhancement-based immunoassay has been developed for the detection of the polycyclic aromatic hydrocarbons (PAH), benzo[a]pyrene (BaP), in aqueous solutions. The results of this study show that BaP, which inefficiently fluoresces in aqueous solution, displays enhanced fluorescence when bound to the anti-BaP antibody (anti-BaP), as part of a label-free immunoassay system. Binding to anti-BaP results in a 3.12-fold increase in the fluorescence intensity of BaP, which emits at 435 nm when excited at 280 nm, due to the hydrophobic interaction and fluorescence resonance energy transfer (FRET) between antibody and antigen. As result of this phenomenon, the antibody-based fluorescence immunoassay system can be used to detect BaP specifically with a limit of detection (LOD) of 0.06 ng mL(-1). Finally, extraction recoveries of BaP from spiked wheat and barley samples were found to be in the range of 80.5-87.0% and 92.9-92.1%, respectively.
已开发出一种基于荧光增强的免疫分析法，用于检测水溶液中的多环芳烃（PAH）——苯并[a]芘（BaP）。本研究结果表明，在水溶液中荧光效率较低的BaP，作为无标记免疫分析系统的一部分，当与抗BaP抗体（anti-BaP）结合时会显示出增强的荧光。由于抗体与抗原之间的疏水相互作用和荧光共振能量转移（FRET），与anti-BaP结合会使BaP的荧光强度增加3.12倍，当在280nm激发时，BaP在435nm处发射荧光。由于这种现象，基于抗体的荧光免疫分析系统可用于特异性检测BaP，检测限（LOD）为0.06 ng mL(-1)。最后，加标小麦和大麦样品中BaP的提取回收率分别为80.5 - 87.0%和92.9 - 92.1%。
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