Rao Lanka Tata, Raz Adva, Patolsky Fernando
Center for Nanoscience and Nanotechnology, Tel Aviv University, Tel Aviv-Yafo, Israel.
School of Chemistry, Faculty of Exact Sciences, Tel Aviv University, Tel Aviv-Yafo, Israel.
Nat Protoc. 2025 Jan 30. doi: 10.1038/s41596-024-01124-6.
Nanostructured devices have proven useful in a broad range of applications, from diagnosing diseases to discovering and screening new drug molecules. We developed vertical silicon nanopillar (SiNP) arrays for on-chip multiplex capture of selected biomolecules using a light-induced release of the array's selectively captured biomarkers. This platform allows the rapid, reusable and quantitative capture and release of a selection of biomarkers, followed by their downstream analysis. Here we outline a standardized protocol for producing the SiNP-based capture-and-release device, which involves the detailed fabrication steps for single-zone nanopillar arrays, their morphological characterization and the chemical modification procedures applied for the anchoring of selective bioreceptors together with the light-controlled on-demand release of the chemical agent. In addition, we provide a detailed approach for the fabrication of a multizone-SiNP array, allowing the simultaneous capture and release of multiple biomarkers of interest. Finally, we demonstrate the entire process of selective and quantitative capture and release of biomolecules from biosamples by means of a commercial low-volume microplate reader system, using green fluorescent protein as a biomarker example. The entire protocol can be conducted within 45 h and requires knowledge in nanoscience, surface chemistry, device micro- and nanofabrication procedures, microfluidics and protein quantification techniques. These SiNP array devices have already demonstrated applications for highly selective and quantitative analysis of a wide range of biological and chemical species, including proteins, nucleic acids, small molecules and ionic species.
纳米结构器件已在广泛的应用中证明了其有用性，从疾病诊断到新药分子的发现和筛选。我们开发了垂直硅纳米柱（SiNP）阵列，用于通过光诱导释放阵列选择性捕获的生物标志物来在芯片上多重捕获选定的生物分子。该平台允许快速、可重复使用且定量地捕获和释放一系列生物标志物，随后进行下游分析。在此，我们概述了一种用于生产基于SiNP的捕获和释放装置的标准化方案，其中涉及单区纳米柱阵列的详细制造步骤、其形态表征以及用于固定选择性生物受体的化学修饰程序，以及化学试剂的光控按需释放。此外，我们提供了一种制造多区SiNP阵列的详细方法，允许同时捕获和释放多种感兴趣的生物标志物。最后，我们以绿色荧光蛋白作为生物标志物示例，通过商用小体积微孔板读取器系统展示了从生物样品中选择性和定量捕获及释放生物分子的整个过程。整个方案可在45小时内完成，并且需要纳米科学、表面化学、器件微纳制造程序、微流体和蛋白质定量技术方面的知识。这些SiNP阵列器件已经在对包括蛋白质、核酸、小分子和离子物种在内的广泛生物和化学物种的高选择性和定量分析中展示了应用。
