Laboratory of Chemical Biology, Division of Biological Inorganic Chemistry, State Key Laboratory of Rare Earth, Resource Utilization, Changchun Institute of Applied Chemistry, Graduate School of the Chinese, Academy of Sciences, Chinese Academy of Sciences, Changchun, Jilin, 130022, China.
Small. 2014 Jan 29;10(2):330-6. doi: 10.1002/smll.201301273. Epub 2013 Aug 5.
Upconversion nanoparticles (UCNPs) have been proposed as a promising new class of biological luminescent labels because of their weak auto-fluorescence background, strong penetration ability under near-infrared (NIR) radiation, resistance to photobleaching, and low toxicity. Although UCNPs hold great promise in nanotechnology and nanomedicine, their applications in ECL fields still remain unexplored. Herein, a label-free, ultra-sensitive and selective electrochemiluminescence (ECL) assay is developed for detection of cyclin A2 by using highly efficient ECL graphene-upconversion hybrid. Being an important member of the cyclin family, cyclin A2 is involved in the initiation of DNA replication, transcription and cell cycle reg-ulation through the association of cyclin-dependent kinases (CDK). Cyclin A2 is a prognostic indicator in early-stage cancers and a target for treatment of different types of cancers. However, the expression level of cyclin A2 is quite low, direct detection of cyclin A2 in crude cancer cell extracts is challenging and important for both clinical diagnosis of cancer in the early stage and the treatment. By chemically grafting cyclin A2 detection specific probe, a PEGlyted hexapeptide, to graphene-upconversion hybrid, the constructed ECL biosensor displays a superior performance for cyclin A2 , which can not only detect cyclin A2 directly in cancer cell extracts, but also discriminate between normal cells and cancer cells. More importantly, the ECL biosensor has different responses between clinical used anticancer drug-treated and non-treated cancer cells, which demonstrates that the sensor can be potentially used for drug screening, and for evaluation of therapeutic treatments in early-stage cancers.
上转换纳米粒子（UCNPs）由于其弱的自体荧光背景、在近红外（NIR）辐射下的强穿透能力、抗光漂白性和低毒性，已被提议作为一类有前途的新型生物发光标记物。尽管 UCNPs 在纳米技术和纳米医学中有很大的应用前景，但它们在 ECL 领域的应用仍未被探索。在此，通过使用高效的电致化学发光（ECL）石墨烯-上转换杂化，开发了一种无标记、超灵敏和选择性的 ECL 分析方法，用于检测细胞周期蛋白 A2。作为细胞周期蛋白家族的重要成员，细胞周期蛋白 A2 通过与细胞周期依赖性激酶（CDK）的结合，参与 DNA 复制、转录和细胞周期调节的启动。细胞周期蛋白 A2 是早期癌症的预后指标，也是不同类型癌症治疗的靶点。然而，细胞周期蛋白 A2 的表达水平相当低，直接检测粗提癌细胞提取物中的细胞周期蛋白 A2 具有挑战性，对早期癌症的临床诊断和治疗都很重要。通过将细胞周期蛋白 A2 检测特异性探针，即 PEG 化六肽，化学接枝到石墨烯-上转换杂化上，构建的 ECL 生物传感器对细胞周期蛋白 A2 表现出优异的性能，不仅可以直接检测癌细胞提取物中的细胞周期蛋白 A2，还可以区分正常细胞和癌细胞。更重要的是，该 ECL 生物传感器在临床使用的抗癌药物治疗和未治疗的癌细胞之间有不同的反应，这表明该传感器可潜在用于药物筛选，并用于早期癌症的治疗评估。
