NNL-Institute of Nanoscience-NANO, CNR, Via per Arnesano 16, Lecce I-73100, Italy.
Biosens Bioelectron. 2013 Apr 15;42:646-52. doi: 10.1016/j.bios.2012.10.012. Epub 2012 Oct 22.
Novel synthetic peptides represent smart molecules for antigen-antibody interactions in several bioanalytics applications, from purification to serum screening. Their immobilization onto a solid phase is considered a key point for sensitivity increasing. In this view, we exploited Quartz Crystal Microbalance with simultaneous frequency and dissipation monitoring (QCM-D) with a double aim, specifically, as investigative tool for spacers monolayer assembling and its functional evaluation, as well as high sensitive method for specific immunosorbent assays. The method was applied to pancreatic ductal adenocarcinoma (PDAC) detection by studying the interactions between synthetic phosphorylated and un-phosphorylated α-enolase peptides with sera of healthy and PDAC patients. The synthetic peptides were immobilized on the gold surface of the QCM-D sensor via a self-assembled alkanethiol monolayer. The presented experimental results can be applied to the development of surfaces less sensitive to non-specific interactions with the final target to suggest specific protocols for detecting PDAC markers with un-labeled biosensors.
新型合成肽是几种生物分析应用中抗原-抗体相互作用的智能分子,从纯化到血清筛选。将其固定在固相上被认为是提高灵敏度的关键。为此,我们利用石英晶体微天平(QCM-D)进行同时监测频率和耗散的方法,具有双重目的,即作为研究间隔物单层组装及其功能评估的研究工具,以及用于高灵敏度特定免疫吸附测定的方法。该方法通过研究合成磷酸化和非磷酸化α-烯醇化酶肽与健康和 PDAC 患者血清之间的相互作用,应用于胰腺导管腺癌(PDAC)的检测。合成肽通过自组装的烷硫醇单层固定在 QCM-D 传感器的金表面上。所提出的实验结果可应用于开发对非特异性相互作用不太敏感的表面,最终目标是建议使用未标记的生物传感器检测 PDAC 标志物的具体方案。