School of Chemistry, the Raymond and Beverly Sackler Faculty of Exact Sciences, Tel-Aviv University, Tel Aviv 69978, Israel.
Nano Lett. 2012 Sep 12;12(9):4748-56. doi: 10.1021/nl3021889. Epub 2012 Aug 2.
The development of efficient biomolecular separation and purification techniques is of critical importance in modern genomics, proteomics, and biosensing areas, primarily due to the fact that most biosamples are mixtures of high diversity and complexity. Most of existent techniques lack the capability to rapidly and selectively separate and concentrate specific target proteins from a complex biosample, and are difficult to integrate with lab-on-a-chip sensing devices. Here, we demonstrate the development of an on-chip all-SiNW filtering, selective separation, desalting, and preconcentration platform for the direct analysis of whole blood and other complex biosamples. The separation of required protein analytes from raw biosamples is first performed using a antibody-modified roughness-controlled SiNWs (silicon nanowires) forest of ultralarge binding surface area, followed by the release of target proteins in a controlled liquid media, and their subsequent detection by supersensitive SiNW-based FETs arrays fabricated on the same chip platform. Importantly, this is the first demonstration of an all-NWs device for the whole direct analysis of blood samples on a single chip, able to selectively collect and separate specific low abundant proteins, while easily removing unwanted blood components (proteins, cells) and achieving desalting effects, without the requirement of time-consuming centrifugation steps, the use of desalting or affinity columns. Futhermore, we have demonstrated the use of our nanowire forest-based separation device, integrated in a single platform with downstream SiNW-based sensors arrays, for the real-time ultrasensitive detection of protein biomarkers directly from blood samples. The whole ultrasensitive protein label-free analysis process can be practically performed in less than 10 min.
高效的生物分子分离和纯化技术的发展在现代基因组学、蛋白质组学和生物传感领域至关重要,主要是因为大多数生物样品都是高度多样化和复杂的混合物。现有的大多数技术都缺乏从复杂的生物样品中快速和选择性地分离和浓缩特定目标蛋白质的能力,并且难以与芯片上的传感设备集成。在这里,我们展示了一种用于直接分析全血和其他复杂生物样品的芯片上全-SiNW 过滤、选择性分离、脱盐和预浓缩平台的开发。首先,使用具有超大结合面积的抗体修饰粗糙度控制 SiNWs(硅纳米线)森林,从原始生物样品中分离所需的蛋白质分析物,然后在受控的液体介质中释放目标蛋白质,并在同一芯片平台上制造的超灵敏 SiNW 基 FET 阵列中对其进行后续检测。重要的是,这是首次在单个芯片上展示用于全血直接分析的全-NWs 设备,能够选择性地收集和分离特定的低丰度蛋白质,同时轻松去除不需要的血液成分(蛋白质、细胞)并实现脱盐效果,而无需耗时的离心步骤、脱盐或亲和柱的使用。此外,我们已经证明了我们的基于纳米线森林的分离装置的用途,该装置集成在单个平台上,与下游基于 SiNW 的传感器阵列结合使用,可直接从血液样本中实时超灵敏地检测蛋白质生物标志物。整个超灵敏蛋白质无标记分析过程可在不到 10 分钟的时间内实际完成。