Development of a cantilever sensor chip as a point-of-care (POC) device for label-free, rapid, and multiplexed detection of cancer protein biomarkers

开发悬臂传感器芯片作为即时护理 (POC) 设备，用于癌症蛋白生物标志物的无标记、快速和多重检测

• 批准号: 244492497
• 负责人: Professor Dr.-Ing. Gerald A. Urban, since 6/2014
• 金额: --
• 依托单位: Institut für Mikrosystemtechnik (IMTEK)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/244492497?language=en
• 关键词: Development; cantilever; sensor; chip; point

High-throughput identification and quantitative detection of disease-specific protein biomarkers forms the foundation of many diagnostic tests to direct therapy in diverse area of clinical medicine such as cancer. Various protein detection platforms have been developed to provide a multi-analytes, specific, and sensitive assay; among them the enzyme-linked immunosorbent assay (ELISA) is a golden standard. While the label-based techniques are widely utilized in those platforms for signal transduction, the label incorporation itself can be highly heterogeneous and the labeling procedure is laborious, lengthy, and adds significant cost and time for development. Therefore, there is great interest in developing label-free techniques for biological analysis. Micro/nano-cantilever-based sensors have been considered as one of the most promising label-free techniques because of the simple and efficient sensing mechanism by translating molecular interactions into mechanics. By utilizing a change in contact angle of a liquid meniscus on a solid surface through molecular interactions such as antigen-antibody binding, we could show that a tiny molecular interaction force is converted to a capillary force-a much larger physical quantity. This physical phenomenon could be served not only as a new mechanism for label-free protein detection but also as a practical method for a sensing platform design. We have previously demonstrated a cantilever sensor design by utilizing such a sensing mechanism for a label-free protein immunoassay with detection limit as low as 1 pg/mL and a short detection time within 10 minutes. In this project we will further investigate (i) new surface modification strategies for efficient molecular recognition as well as surface passivation, (ii) new cantilever design and fabrication process to improve the signal-to-noise ratio and linear dynamic range of sensor, and (iii) integrated microfluidics for sample processing on a single sensor chip for label-free, rapid, and multiplexed detection of cancer protein biomarkers. The proposed sensor chip design leads to the construction of an easy-to-use immunosensing platform intended for facile handling, minimal sample/reagent requirements and compatibility with diverse biomarkers. Biological samples from cancer patients are planned to be applied to the sensor chip to detect a panel of 5 cancer protein biomarkers, in an effort to provide a point-of-care (POC) test alternative to current centralized testing for biomarkers.

疾病特异性蛋白生物标志物的高通量鉴定和定量检测形成了许多诊断试验的基础，以指导癌症等临床医学的不同领域的治疗。各种蛋白质检测平台已经被开发出来，以提供多分析、特异和灵敏的分析，其中，酶联免疫吸附试验(ELISA)是金标准。虽然基于标记的技术被广泛应用于这些信号转导平台中，但标记掺入本身可能是高度异质性的，标记过程费时费力，并增加了开发的大量成本和时间。因此，发展生物分析的无标记技术引起了人们的极大兴趣。微/纳米悬臂梁传感器通过将分子间的相互作用转化为力学机制，具有简单高效的传感机理，被认为是最有前途的无标记传感技术之一。通过利用液体半月板在固体表面上通过分子相互作用(如抗原-抗体结合)接触角的变化，我们可以证明微小的分子相互作用力被转化为毛细管力--一个大得多的物理量。这一物理现象不仅可以作为无标记蛋白质检测的新机制，也可以作为传感平台设计的实用方法。我们之前已经展示了悬臂式传感器的设计，利用这种传感机制进行了无标记蛋白质免疫分析，检测下限低至1pg/mL，检测时间在10分钟内很短。在这个项目中，我们将进一步研究(I)用于高效分子识别和表面钝化的新的表面修饰策略，(Ii)用于提高传感器的信噪比和线性动态范围的新的悬臂梁设计和制造工艺，以及(Iii)用于在单个传感器芯片上进行样品处理的集成微流控技术，以实现对癌症蛋白质生物标志物的无标记、快速和多路检测。建议的传感器芯片设计导致构建一个易于使用的免疫传感平台，旨在方便处理，最大限度地减少样品/试剂需求，并与不同的生物标记物兼容。来自癌症患者的生物样本计划应用于传感器芯片，以检测由5个癌症蛋白质生物标记物组成的面板，以努力提供一种替代目前集中检测生物标记物的护理点(POC)测试。