title: Multiple signal amplification electrogenerated chemiluminescence biosensors for sensitive protein kinase activity analysis and inhibition.
abstract: 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.
authors: 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.
authors: Zonghua
authors: Z
authors: Wang
authors: 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.
authors: Zhiyong
authors: Z
authors: Yan
authors: 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.
authors: Na
authors: N
authors: Sun
authors: Department of Chemistry, Beijing Key Laboratory for Analytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China. Electronic address: liu-yang@mail.tsinghua.edu.cn.
authors: Yang
authors: Y
authors: Liu
mesh_terms: D015374:Biosensing Techniques; D004247:DNA / Q000737:chemistry; D006046:Gold; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D019271:Hypoxanthine / Q000737:chemistry; D057230:Limit of Detection; D008165:Luminol / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry*; D047428:Protein Kinase Inhibitors / Q000737:chemistry; D011494:Protein Kinases / Q000737:chemistry / Q000302:isolation & purification*; D014969:Xanthine Oxidase / Q000737:chemistry
chemical_list: D047428:Protein Kinase Inhibitors; D019271:Hypoxanthine; D008165:Luminol; D006046:Gold; D004247:DNA; D006861:Hydrogen Peroxide; D014969:Xanthine Oxidase; D011494:Protein Kinases
keywords: Electrogenerated chemiluminescence; Enzyme; Gold nanoparticle; Kinase; Phosphorylation