Integrated Microfluidic Aptasensor for Mass Spectrometric Detection of Vasopressin in Human Plasma Ultrafiltrate.
Integrated Microfluidic Aptasensor for Mass Spectrometric Detection of Vasopressin in Human Plasma Ultrafiltrate.
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DOI：
10.1039/c5ay02979a
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发表时间：
2016-07-14
期刊：
影响因子：
--
通讯作者：
Lin Q
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文献类型：
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作者：
Yang J;Zhu J;Pei R;Oliver JA;Landry DW;Stojanovic MN;Lin Q
我们提出了一种基于微流控适体的生物传感器，用于检测患者样本中的低分子量生物标志物。使用微流控装置，集成了基于适体的特定分析物提取，等度洗脱，并通过基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱检测，我们展示了快速，灵敏和无标记检测精氨酸加压素（AVP）在人血浆超滤液。复杂基质中的AVP分子被通过微室中的亲和结合固定在微珠上的适体特异性捕获。在通过洗涤去除未结合的污染分子后，通过芯片上温度控制热破坏适体-AVP复合物。释放的AVP分子用纯化水洗脱并转移到单独的微室中，并通过重复的压电致动喷射沉积到MALDI板上的单个点上，这在点区域上富集AVP分子。这种集成的芯片上样品处理能够通过MALDI-TOF质谱法以快速和无标记的方式定量检测低丰度AVP。我们的实验结果表明，通过基于适体的选择性预浓缩，在人血浆超滤液中检测AVP低至生理相关的皮摩尔浓度，证明了我们的方法作为快速（约1小时），灵敏的临床AVP测定的潜力。
We present a microfluidic aptamer-based biosensor for detection of low-molecular-weight biomarkers in patient samples. Using a microfluidic device that integrates aptamer-based specific analyte extraction, isocratic elution, and detection by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, we demonstrate rapid, sensitive and label-free detection of arginine vasopressin (AVP) in human plasma ultrafiltrate. AVP molecules in complex matrices are specifically captured by an aptamer that is immobilized on microbeads via affinity binding in a microchamber. After the removal of unbound, contaminating molecules through washing, aptamer-AVP complexes are thermally disrupted via on-chip temperature control. Released AVP molecules are eluted with purified water and transferred to a separate microchamber, and deposited onto a single spot on a MALDI plate via repeated, piezoelectrically actuated ejection, which enriches AVP molecules over the spot area. This integrated on-chip sample processing enables the quantitative detection of low-abundance AVP by MALDI-TOF mass spectrometry in a rapid and label-free manner. Our experimental results show the detection of AVP in human plasma ultrafiltrate as low as physiologically relevant picomolar concentrations via aptamer-based selective preconcentration, demonstrating the potential of our approach as a rapid (~ 1hr), sensitive clinical AVP assay.