SPEI Biosensor Development and Optimization

SPEI生物传感器的开发和优化

• 批准号: 7096425
• 负责人: DALE NORMAN LARSON
• 金额: $ 25.09万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7096425
• 关键词: antibody; clinical research; lighting; microfluidics; oligonucleotides; peptides; performance; surface plasmon resonance

DESCRIPTION (provided by applicant): Biosensors are important tools for both research and clinical diagnostics. Since the range of applications for biosensors is broad and the requirements varied, the ideal biosensor technology would combine sensitivity with facile multiplexing and enable the detection and quantification of analytes without the need for a label. It would also be compatible with many different capture agents (e.g. antibodies, aptamers, peptides) and provide real-time detection for the collection of kinetic, as well as equilibrium, binding data. Currently available biosensors do not meet all of these demanding criteria. Here, we propose that a biosensor based on surface plasmon enhanced illumination (SPEI) addresses these issues and thus is expected to have a broad impact in the areas of basic biology, translational research, and clinical diagnostics. The SPEI biosensor that we describe in this proposal is based on measuring changes in the amount of light transmitted through nanoscale (50-200nm) apertures in a metallic surface. We and others have found that the amount of transmitted light depends strongly on the local index of refraction at the sensor surface, which is altered upon the binding of analytes to immobilized capture reagents. While biosensors based on surface plasmon resonance (SPR) also detect changes in the local index of refraction due to biomolecular interactions, the plasmon excitation method is very different for-the SPEI technology. These differences, described in detail in this proposal, allow the active sensing area to be very small, enabling a very high degree of multiplexing, superior resolution, and an increased sensitivity of at least 20 fold over grating coupled SPR. The similarity to conventional SPR, however, allows for the body of applications work done for conventional SPR to be used with SPEI. We propose to design and fabricate a multiplexed SPEI biosensor and demonstrate its performance by detecting and quantifying multiple cytokines in complex media. The specific issues that will be addressed include developing appropriate surface chemistry to reduce nonspecific binding, developing methods that enable the accurate and simultaneous quantification of multiple analytes, and developing methods to optimize the specificity of detection (reduce the incidence of false positives).

描述（由申请人提供）：生物传感器是研究和临床诊断的重要工具。由于生物传感器的应用范围很广，并且需求各不相同，因此理想的生物传感器技术将敏感性与便捷的多路复用相结合，并可以在不需要标签的情况下对分析物进行检测和量化。它也将与许多不同的捕获剂（例如抗体，适体，肽）兼容，并为收集动力学以及平衡，结合数据提供实时检测。目前可用的生物传感器不符合所有这些苛刻的标准。在这里，我们建议基于表面等离子体增强照明（SPEI）的生物传感器解决这些问题，因此有望在基本生物学，翻译研究和临床诊断方面产生广泛的影响。我们在该提案中描述的SPEI生物传感器是基于测量金属表面中通过纳米级（50-200nm）光圈传输的光量的变化。我们和其他人发现，传输光的量强烈取决于传感器表面的局部折射指数，这在分析物与固定捕获试剂的结合后会改变。尽管基于表面等离子体共振（SPR）的生物传感器也检测到由于生物分子相互作用而引起的局部折射指数的变化，但对于SPEI技术，等离子体激发方法却大不相同。这些差异在此提案中详细描述，使主动传感区域非常小，从而实现了非常高的多重程度，优质的分辨率，并且在光栅耦合的SPR上提高了至少20倍的灵敏度。然而，与常规SPR的相似性允许使用SPEI使用用于常规SPR的应用程序。我们建议设计和制造多重SPEI生物传感器，并通过检测和量化复杂培养基中的多个细胞因子来证明其性能。将要解决的具体问题包括开发适当的表面化学以减少非特异性结合，开发能够准确和同时量化多个分析物的方法，以及开发方法以优化检测的特异性（降低误报的发生率）。