Institute of Photonics and Electronics, AS CR Chaberská 57, 182 51 Prague, Czech Republic.
Anal Chem. 2013 Jun 18;85(12):5637-40. doi: 10.1021/ac401062m. Epub 2013 May 28.
We present a novel approach to reference-compensated label-free affinity biosensing in complex media. Unlike conventional approaches that employ surfaces with different biological functionalities in the detection and reference channels to produce a reference-compensated sensor response, the new approach (referred as to single surface referencing (SSR)) uses a single functionalized surface split into the detection and reference channel to which complex sample (detection channel) and complex sample mixed with biomolecules binding to the analyte and thus inhibiting the binding of the analyte to the functionalized surface (reference channel) is introduced. This approach ensures that (i) only the detection channel captures the analyte and (ii) nonspecific binding incurred in the detection and reference channels are the same. We evaluate this approach in a model biosensing experiment, detection of a cancer biomarker carcinoembryonic antigen (CEA) in blood plasma using antibody against CEA and a surface plasmon resonance (SPR) biosensor. We detect CEA in three different blood plasma samples and demonstrate that this novel referencing approach provides more accurate results and lower biological variability than the conventional referencing.
我们提出了一种新的方法来在复杂介质中进行参考补偿的无标记亲和生物传感。与传统方法不同，传统方法在检测和参考通道中使用具有不同生物学功能的表面来产生参考补偿的传感器响应，而新方法（称为单表面参考（SSR））使用分为检测和参考通道的单个功能化表面，其中引入了复杂样品（检测通道）和与分析物结合并因此抑制分析物与功能化表面结合的生物分子混合物（参考通道）。这种方法确保 (i) 仅检测通道捕获分析物，以及 (ii) 在检测和参考通道中发生的非特异性结合是相同的。我们在模型生物传感实验中评估了这种方法，即使用针对癌胚抗原 (CEA) 的抗体和表面等离子体共振 (SPR) 生物传感器检测血浆中的癌症生物标志物 CEA。我们在三种不同的血浆样本中检测到 CEA，并证明这种新的参考方法比传统参考方法提供更准确的结果和更低的生物学变异性。
