用于灵敏蛋白激酶活性分析和抑制的多重信号放大电化学发光生物传感器。
Multiple signal amplification electrogenerated chemiluminescence biosensors for sensitive protein kinase activity analysis and inhibition.
机构信息
Collaborative Innovation Center for Marine Biomass Fiber Materials and Textiles, College of Chemical Science and Engineering, Shandong Sino-Japanese Center for Collaborative Research of Carbon Nanomaterials, Laboratory of Fiber Materials and Modern Textiles, The Growing Base for State Key Laboratory, Qingdao University, Qingdao, Shandong 266071, China.
Department of Chemistry, Beijing Key Laboratory for Analytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China; Collaborative Innovation Center for Marine Biomass Fiber Materials and Textiles, College of Chemical Science and Engineering, Shandong Sino-Japanese Center for Collaborative Research of Carbon Nanomaterials, Laboratory of Fiber Materials and Modern Textiles, The Growing Base for State Key Laboratory, Qingdao University, Qingdao, Shandong 266071, China.
出版信息
Biosens Bioelectron. 2015 Jun 15;68:771-776. doi: 10.1016/j.bios.2015.02.006. Epub 2015 Feb 7.
A novel electrogenerated chemiluminescence (ECL) biosensor was built for the detection of kinase activity based on multiple signal amplification nanoprobes. In this strategy, the Xanthine oxidase (XOD) and 5'-phosphate group end DNA conjugated AuNPs was integrated with the phosphorylated peptide by Zr(4+). The XOD on gold nanoparticles can catalyze dissolved oxygen to produce H2O2 in the presence of hypoxanthine (HA) which acts as a coreactor for luminol ECL reaction. In addition, due to the excellent catalytic activity of gold nanoparticle toward luminol ECL reaction and its large surface area that can accommodate large number of XOD and DNA on the surface, the ECL signal of luminol was significantly amplified, affording a highly sensitive ECL analysis of kinase activity. The as-proposed biosensor presents a low detection limit of 0.09 U mL(-1) for protein kinase A (PKA) activity, wide linear range (from 0.1 to 10 U mL(-1)) and excellent stability even in serum samples. This biosensor can also be applied for quantitative kinase inhibitor evaluation. The robust ECL biosensor provides a valuable tool for the high throughput assay in the applications of clinic diagnostic and therapeutic.
一种基于多重信号放大纳米探针的新型电致化学发光（ECL）生物传感器被构建用于检测激酶活性。在该策略中，黄嘌呤氧化酶（XOD）和 5'-磷酸基团末端 DNA 与通过 Zr(4+) 与磷酸化肽结合。在黄嘌呤（HA）存在下，金纳米粒子上的 XOD 可以催化溶解氧产生 H2O2，HA 作为鲁米诺 ECL 反应的核心反应剂。此外，由于金纳米粒子对鲁米诺 ECL 反应具有出色的催化活性，并且其表面积大，可以在表面容纳大量的 XOD 和 DNA，因此鲁米诺的 ECL 信号得到了显著放大，实现了对激酶活性的高灵敏度 ECL 分析。所提出的生物传感器对蛋白激酶 A（PKA）活性的检测限低至 0.09 U mL(-1)，线性范围宽（从 0.1 到 10 U mL(-1))，即使在血清样本中也具有出色的稳定性。该生物传感器还可用于定量激酶抑制剂评估。这种强大的 ECL 生物传感器为临床诊断和治疗中的高通量分析提供了有价值的工具。
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