Biomedical Diagnostics Institute, National Centre for Sensor Research, School of Physical Sciences, Dublin City University, Dublin, Ireland.
Biosens Bioelectron. 2010 Feb 15;25(6):1344-9. doi: 10.1016/j.bios.2009.10.026. Epub 2009 Oct 28.
We have carried out a human IgG immunoassay on a novel disposable optical array biochip using surface plasmon-coupled emission (SPCE) detection. The work successfully combines the advantages of the highly directional SPCE emission profile and enhanced surface plasmon excitation with the high light collection efficiency achieved using supercritical angle fluorescence (SAF). This is achieved using an array of transparent paraboloid polymer elements which have been coated with a thin gold layer to facilitate SPCE. Moreover, since only the emission of molecules which are close to the metal surface couple into the surface plasmon, the detection is highly surface-specific leading to background suppression and increased signal-to-noise ratio. Theoretical calculations have been carried out in order to match the surface plasmon resonance angles and SPCE emission angles to the paraboloid array features for light collection. A sandwich assay format was used and a dose response curve was obtained in the concentration range 2 ng/ml to 200 microg/ml yielding a limit of detection of 20 ng/ml. This is the first demonstration of an SPCE-based assay on a disposable biochip platform and indicates the potential of SPCE-based arrays for high-throughput analysis of biomolecular interactions.
我们使用表面等离子体耦合发射(SPCE)检测,在新型一次性光学生物芯片上进行了人 IgG 免疫分析。该工作成功地结合了高度定向的 SPCE 发射谱和增强的表面等离子体激发的优点,以及使用超临界角荧光(SAF)实现的高光收集效率。这是通过涂覆有薄金层以促进 SPCE 的透明抛物面聚合物元件阵列来实现的。此外,由于只有靠近金属表面的分子的发射才能耦合到表面等离子体中,因此检测具有高度的表面特异性,从而抑制背景并提高信噪比。为了将表面等离子体共振角和 SPCE 发射角与光收集的抛物面阵列特征相匹配,进行了理论计算。采用三明治测定法,在 2ng/ml 至 200μg/ml 的浓度范围内获得了剂量反应曲线,检测限为 20ng/ml。这是在一次性生物芯片平台上进行基于 SPCE 的测定的首次证明,并表明基于 SPCE 的阵列在生物分子相互作用的高通量分析中具有潜力。