Centre for Nanometrology, Department of Pure and Applied Chemistry, WestCHEM, University of Strathclyde, 295 Cathedral Street, Glasgow, UK.
Analyst. 2011 Jul 21;136(14):2925-30. doi: 10.1039/c1an15178a. Epub 2011 Jun 7.
Advancements in lithography methods for printing biomolecules on surfaces are proving to be potentially beneficial for disease screening and biological research. Dip-pen nanolithography (DPN) is a versatile micro and nanofabrication technique that has the ability to produce functional biomolecule arrays. The greatest advantage, with respect to the printing mechanism, is that DPN adheres to the sensitive mild conditions required for biomolecules such as proteins. We have developed an optimised, high-throughput printing technique for fabricating protein arrays using DPN. This study highlights the fabrication of a prostate specific antigen (PSA) immunoassay detectable by fluorescence. Spot sizes are typically no larger than 8 μm in diameter and limits of detection for PSA are comparable with a commercially available ELISA kit. Furthermore, atomic force microscopy (AFM) analysis of the array surface gives great insight into how the nitrocellulose substrate functions to retain protein integrity. This is the first report of protein arrays being printed on nitrocellulose using the DPN technique and the smallest feature size yet to be achieved on this type of surface. This method offers a significant advance in the ability to produce dense protein arrays on nitrocellulose which are suitable for disease screening using standard fluorescence detection.
在表面印刷生物分子的光刻方法的进步被证明对疾病筛查和生物研究具有潜在的益处。蘸笔纳米光刻（DPN）是一种多功能的微纳制造技术，具有制造功能生物分子阵列的能力。就打印机制而言，最大的优势是 DPN 符合蛋白质等生物分子所需的敏感温和条件。我们已经开发了一种优化的、高通量的 DPN 打印技术，用于制造蛋白质阵列。本研究重点介绍了使用荧光法检测前列腺特异性抗原（PSA）免疫测定的制造。斑点大小通常不超过 8μm 直径，PSA 的检测限与市售的 ELISA 试剂盒相当。此外，对阵列表面的原子力显微镜（AFM）分析深入了解了硝化纤维素基质如何保持蛋白质完整性。这是首次报道使用 DPN 技术在硝化纤维素上打印蛋白质阵列，也是迄今为止在这种类型的表面上实现的最小特征尺寸。该方法在在硝化纤维素上制造适合使用标准荧光检测进行疾病筛查的密集蛋白质阵列方面取得了重大进展。
