组成图示
点击图片查看大图 · 依据论文自动绘制
传感器类型
荧光生物传感器
检测对象
多环芳烃（polycyclic aromatic hydrocarbons, PAHs；3-5环PAHs，以phenanthrene equivalents计）；样品基质：河口疏浚区水样、雨水径流水样
检测原理
该传感器基于KinExA竞争免疫荧光原理。PMMA微珠表面包被DBT-AA-BSA抗原，作为固相捕获层。用AlexaFluor 647标记的单克隆抗体7B2.3与固相抗原结合；样品中3-5环PAHs与标记抗体竞争结合抗原。PAH浓度越高，结合到微珠上的荧光抗体越少，620 nm激发、670 nm发射的荧光信号越低。荧光信号与PAH浓度呈对数线性负相关。抗体-抗原平衡约10 s，无需额外孵育，定量约3 min。使用phenanthrene标准建立校准曲线，结果以phenanthrene equivalents报告。
检测灵敏度
LOD: 0.3 mg/L (phenanthrene equivalents, EPA method)；线性范围: 0.3 to 30 mg/L；field LOD: 0.2 to 1 mg/L；R^2 ≈ 0.98
效应效果
该传感器现场快速、便携，定量响应约3 min，采样后10 min内出结果，3 d内完成80+样品。标准品重复误差通常≤10%。与GC-MS比较：河口疏浚区水样r=0.95（n=7），Deming斜率0.90±0.14；雨水径流水样r=0.86（n=7），斜率9.68±2.63。空间监测定义0.3–3.2 mg/L（phenanthrene equivalents）羽流，覆盖约90,000 m2，确认PAH未超10 mg/L；降雨峰值4.4和3.6 mg/L。抗体识别3–5环PAHs，交叉反应可累加低水平烷基化PAHs。作者认为其可指导采样与修复决策，节省时间和成本。
传感器的构成
- 检测平台：KinExA Inline Sensor flow cell，容纳微珠并实现荧光检测
- 固相载体：PMMA beads (Sapidyne)，承载抗原包被层
- 固相捕获层：dibenzothiophene-acrylic acid-bovine serum albumin (DBT-AA-BSA)，包被于PMMA微珠，用于结合荧光标记抗体
- 识别元件：monoclonal antibody (mAb) 7B2.3，特异性识别3-5环PAHs
- 信号标记物：AlexaFluor 647标记的mAb 7B2.3，提供荧光信号
- 清洗再生液：50% dimethylsulfoxide (DMSO)/去离子水，样品间清洗管路防止交叉污染
- 校准标准：phenanthrene/PAH标准溶液，用于建立定量校准曲线
中文摘要
本研究展示了利用抗体基生物传感器对水环境中溶解性多环芳烃（PAHs）进行快速、现场、定量评估的两个现场应用。该平台采用 Sapidyne Instruments 的 KinExA 在线传感器，使用对3至5环PAHs具有特异性的单克隆抗体7B2.3。研究在受污染沉积物疏浚作业附近开展空间监测，并在降雨事件期间开展时间监测。最重要的是，近实时数据生成为现场管理决策提供了依据，并确定了用于常规实验室分析的适当采样方案。该方法可在样品采集后10分钟内测定低至0.3 mg/L的PAH浓度，并在3天内完成80个以上样品（不含标准品和空白）的评估。结果与实验室气相色谱-质谱法进行了比较，后者检测了包括烷基化同系物在内的多种PAHs。该系统作为快速监测PAH污染的现场仪器展现出良好前景。
英文摘要
Rapid, on-site, quantitative assessments of dissolved polycyclic aromatic hydrocarbons (PAHs) were demonstrated for two field applications. The platform, a KinExA Inline Sensor (Sapidyne Instruments), employed the monoclonal anti-PAH antibody, 7B2.3, which has specificity for 3- to 5-ring PAHs. A spatial study was conducted near a dredging site where contaminated sediments were being removed, and a temporal study was performed during a rainfall event. Most importantly, the generation of near real-time data guided management decisions in the field and determined proper sampling protocols for conventional analyses. The method was able to determine PAH concentrations as low as 0.3 µg/L, within 10 min of sample acquisition, and to assess 80+ samples (not including standards and blanks) in less than 3 d. These results were compared with a laboratory-based gas chromatography-mass spectrometry method in which a wide array of PAHs, including alkylated homologs, were examined. This system shows great promise as a field instrument for the rapid monitoring of PAH pollution.