文献类型：期刊文献

英文题名：A two-dimensional fluorescence and chemiluminescence orthogonal probe for discriminating and quantifying similar proteins

作者：Li, Juan[1];Zhao, Xiuyan[1];Zhang, Yutao[1];Lu, Yao[1];Xue, Haoyun[1];Li, Dan[1];Liu, Qiang[1];Yan, Chenxu[1];Chi, Weijie[2];Xiao, Xingqing[2];Zhu, Wei-Hong[1];Guo, Zhiqian[1]

机构：[1]East China Univ Sci & Technol, Inst Fine Chem, Feringa Nobel Prize Sci Joint Res Ctr,Frontiers Sc, Sch Chem & Mol Engn,Ctr Photosensit Chem Engn, Shanghai 200237, Peoples R China;[2]Hainan Univ, Sch Chem & Chem Engn, Dept Chem, Haikou 570228, Hainan, Peoples R China

年份：2025

卷号：16

期号：7

起止页码：3228

外文期刊名：CHEMICAL SCIENCE

收录：;EI(收录号：20250417754779);WOS:【SCI-EXPANDED(收录号:WOS:001399308400001)】；

基金：This work was supported by the National Key Research and Development Program (2023YFA1802000), NSFC/China (22225805, 22308101, 22268016, 32394001, and 32121005), Shanghai Science and Technology Innovation Action Plan (No. 23J21901600), Shanghai Frontier Science Research Base of Optogenetic Techniques for Cell Metabolism (Shanghai Municipal Education Commission, grant 2021 Sci & Tech 03-28).

语种：英文

外文关键词：Cycloaddition - Transient absorption spectroscopy

摘要：Given that proteins with minor variations in amino acid sequences cause distinct functional outcomes, identifying and quantifying similar proteins is crucial, but remains a long-standing challenge. Herein, we present a two-dimensional orthogonal fluorescence and chemiluminescence design strategy for the probe DCM-SA, which is sequentially activated by albumin-mediated hydrolysis, exhibiting light-up fluorescence and photo-induced cycloaddition generating chemiluminescence, enabling orthogonal signal amplification for discrimination of subtle differences between similar proteins. By orthogonalizing these dual-mode signals, a two-dimensional work curve of fluorescence and chemiluminescence is established to distinguish and quantify similar proteins HSA and BSA. Importantly, the dual-mode signals of DCM-SA exhibit contrary incremental trends towards HSA and BSA. Molecular docking and femtosecond transient absorbance spectroscopy reveal that the lower KD value of DCM-SA with HSA and the longer excited-state lifetime of DCM-SA with BSA underlie the distinct dual-mode responses. Using two-dimensional orthogonal signals, for the first time, we precisely measure the HSA/BSA ratio in mixed serum. This method facilitates rapid blood source identification and trace HSA quantitation in human urine. Our two-dimensional orthogonal amplification approach offers a powerful tool for distinguishing and quantifying subtle differences among highly similar proteins, demonstrating great potential for both basic life science research and clinical applications.