DNA-directed protein immobilization on mixed self-assembled monolayers via a Streptavidin bridge
DNA-directed protein immobilization on mixed self-assembled monolayers via a Streptavidin bridge
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DOI：
10.1021/la049867r
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发表时间：
2004-09-14
期刊：
影响因子：
3.9
通讯作者：
Jiang, S
中科院分区：
文献类型：
--
作者：
Ladd, J;Boozer, C;Jiang, S
同时检测多种分析物是生物传感器技术进步的重要考虑因素。目前，很少有传感器系统具备准确、精确地检测多种抗原的能力。这项工作提出了一种适合多通道检测的传感器表面功能化的简单方法。该方法利用自组装单层 (SAM) 化学来创建一个无污染的功能性传感器平台，该平台基于通过链霉亲和素桥固定到生物素化链烷硫醇和寡聚乙二醇的混合 SAM 上的生物素化单链 DNA。传感器表面的防污背景可最大限度地减少非特异性结合。通过应用蛋白质-DNA 缀合物生成可用的蛋白质芯片，该缀合物利用 DNA 杂交的特异性定向到传感器芯片表面上的特定位点。所描述的平台在定制的表面等离子共振生物传感器中得到演示。使用该蛋白质阵列的传感器的检测能力已使用人绒毛膜促性腺激素 (hCG) 进行了表征。该平台检测 hCG 的灵敏度高于使用生物素化抗体观察到的灵敏度。结果还显示蛋白质固定到阵列中适当位置的优异特异性。大量可能的 DNA 序列与碱基配对的选择性相结合，使该平台成为能够进行多通道蛋白质检测的传感器的绝佳候选者。
The simultaneous detection of multiple analytes is an important consideration for the advancement of biosensor technology. Currently, few sensor systems possess the capability to accurately and precisely detect multiple antigens. This work presents a simple approach for the functionalization of sensor surfaces suitable for multichannel detection. This approach utilizes self-assembled monolayer (SAM) chemistry to create a nonfouling, functional sensor platform based on biotinylated single-stranded DNA immobilized via a streptavidin bridge to a mixed SAM of biotinylated alkanethiol and oligo(ethylene glycol). Nonspecific binding is minimized with the nonfouling background of the sensor surface. A usable protein chip is generated by applying protein-DNA conjugates which are directed to specific sites on the sensor chip surface by utilizing the specificity of DNA hybridization. The described platform is demonstrated in a custom-built surface plasmon resonance biosensor. The detection capabilities of a sensor using this protein array have been characterized using human chorionic gonadotropin (hCG). The platform shows a higher sensitivity in detection of hCG than that observed using biotinylated antibodies. Results also show excellent specificity in protein immobilization to the proper locations in the array. The vast number of possible DNA sequences combine with the selectivity of base-pairing makes this platform an excellent candidate for a sensor capable of multichannel protein detection.