Feng Duan, Tian Fang, Qin Weijie, Qian Xiaohong
National Center for Protein Sciences Beijing , State Key Laboratory of Proteomics , Beijing Proteome Research Center , Beijing Institute of Radiation Medicine , Beijing , China 102206 . Email:
Chem Sci. 2016 Mar 1;7(3):2246-2250. doi: 10.1039/c5sc03363b. Epub 2015 Dec 15.
Real-time imaging and quantitative monitoring of dynamic changes in biomolecules in bioprocesses is in urgent need for biological and clinical studies. However, this goal is difficult to achieve due to the incompatibility between the commonly used imaging and quantification methods. To fulfill this need, we developed the first nanoprobe capable of both living cell imaging and ultrasensitive quantification of proteases in cancer cells by taking advantage of the characteristic luminescence and ICP-MS response of lanthanide metals. The nanoprobe is composed of a lanthanide metal-based luminescent donor/mass tag and a gold nanoparticle quencher linked by a specific peptide recognized by proteases. The protease activity in living cells is both visualized and quantified by monitoring the enzymatically released lanthanide metal using fluorescence imaging and ICP-MS. The low ppb level sensitivity of this method demonstrates its potential in the study of protease-dependent pathways and related diseases.
生物过程中生物分子动态变化的实时成像和定量监测对于生物学和临床研究而言迫在眉睫。然而,由于常用成像和定量方法之间的不兼容性,这一目标难以实现。为满足这一需求,我们开发了首个纳米探针,它利用镧系金属的特征发光和ICP-MS响应,能够对癌细胞中的蛋白酶进行活细胞成像和超灵敏定量。该纳米探针由镧系金属基发光供体/质量标签和通过蛋白酶识别的特定肽连接的金纳米颗粒猝灭剂组成。通过使用荧光成像和ICP-MS监测酶促释放的镧系金属,可对活细胞中的蛋白酶活性进行可视化和定量。该方法低至ppb级的灵敏度证明了其在蛋白酶依赖性途径及相关疾病研究中的潜力。