Size-selective concentration and label-free characterization of protein aggregates using a Raman active nanofluidic device
Size-selective concentration and label-free characterization of protein aggregates using a Raman active nanofluidic device
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DOI：
10.1039/c0lc00383b
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发表时间：
2011-01-01
期刊：
影响因子：
6.1
通讯作者：
Erickson, David
中科院分区：
文献类型：
--
作者：
Choi, Inhee;Huh, Yun Suk;Erickson, David
蛋白质聚集体的痕量检测和物理化学特征对许多疾病的认识和诊断有很大影响，如衰老相关的神经退行性变和系统性淀粉样变性，蛋白质聚集体的形成是其病理特征之一。在这里，我们展示了一种创新的无标记方法，用于使用拉曼活性纳米流体设备检测和表征少量早期蛋白质聚集体。通过一种新的弹性体坍塌技术形成的亚微米通道使成熟的蛋白质聚集体能够从小的蛋白质分子中分离和浓缩。通过固定在微/纳米流体连接下方的金纳米粒子簇的拉曼增强，可以在比传统分析工具所能完成的低得多的浓度水平(类似于FM)来表征蛋白质聚集体的固有性质。利用我们的设备，我们首次展示了在这些低浓度范围内蛋白质聚集的浓度依赖性。我们期望我们的方法可以促进疾病的早期明确诊断和可能的治疗。
Trace detection and physicochemical characterization of protein aggregates have a large impact in understanding and diagnosing many diseases, such as ageing-related neurodegeneration and systemic amyloidosis, for which the formation of protein aggregates is one of the pathological hallmarks. Here we demonstrate an innovative label-free method for detecting and characterizing small amounts of early stage protein aggregates using a Raman active nanofluidic device. Sub-micrometre channels formed by a novel elastomeric collapse technique enable the separation and concentration of matured protein aggregates from small protein molecules. The Raman enhancement by gold nanoparticle clusters fixed below a micro/nanofluidic junction allows characterization of intrinsic properties of protein aggregates at concentration levels (similar to fM) much lower than can be done with traditional analytical tools. With our device we show for the first time the concentration dependence of protein aggregation over these low concentration ranges. We expect that our method could facilitate definitive diagnosis and possible therapeutics of diseases at early stages.