Yao Chunyan, Qi Yongzhi, Zhao Yuhui, Xiang Yang, Chen Qinghai, Fu Weiling
Department of Laboratory Medicine, Southwest Hospital, Third Military Medical University, Chong Qing 400038, PR China. yao
Biosens Bioelectron. 2009 Apr 15;24(8):2499-503. doi: 10.1016/j.bios.2008.12.036. Epub 2009 Jan 4.
The aim of this study was to develop a rapid method to measure IgE in human serum by use of a direct aptamer-based biosensor based on a quartz crystal microbalance (QCM). An avidin monolayer was applied to immobilize aptamers specific for IgE on the gold surface of a quartz crystal. The frequency shifts (FS) of the QCM were measured and related to IgE concentrations. We could demonstrate that aptamers were able to detect IgE with high specificity and sensitivity in 15 min. A linear relationship existed between the FS (Hz) and the IgE concentrations from 2.5 to 200 microg/L in buffer and human serum. The regression equation was y = 1.03x - 0.06 for this QCM method and chemiluminescence in 50 clinical human serum samples. In addition, the aptamer receptors tolerated repeated affine layer regeneration after ligand binding and recycling of the biosensor with little loss of sensitivity. When stored for 3 weeks, the FS were all greater than 90% of those on the response at the first day. The QCM biosensor can measure IgE and offer advantages of high specificity, reusability, low detection limit, no label or sample pretreatment, and low sample requirement. The aptamer QCM biosensor was suitable for sensitive and specific protein detection, representing an innovative tool for future proteomics.
本研究的目的是开发一种快速方法,通过使用基于石英晶体微天平(QCM)的直接适配体生物传感器来测量人血清中的IgE。将抗生物素蛋白单层应用于在石英晶体的金表面固定对IgE具有特异性的适配体。测量QCM的频率偏移(FS)并将其与IgE浓度相关联。我们能够证明适配体能够在15分钟内以高特异性和灵敏度检测IgE。在缓冲液和人血清中,FS(Hz)与2.5至200μg/L的IgE浓度之间存在线性关系。对于该QCM方法和50份临床人血清样品中的化学发光,回归方程为y = 1.03x - 0.06。此外,适配体受体在配体结合和生物传感器循环后耐受重复的亲和层再生,而灵敏度几乎没有损失。储存3周后,FS均大于第一天响应时的90%。QCM生物传感器可以测量IgE,并具有高特异性、可重复使用性、低检测限、无需标记或样品预处理以及低样品需求等优点。适配体QCM生物传感器适用于灵敏且特异的蛋白质检测,是未来蛋白质组学的一种创新工具。