title: Binding-regulated click ligation for selective detection of proteins.
abstract: Herein, a binding-regulated click ligation (BRCL) strategy for endowing selective detection of proteins is developed with the incorporation of small-molecule ligand and clickable DNA probes. The fundamental principle underlying the strategy is the regulating capability of specific protein-ligand binding against the ligation between clickable DNA probes, which could efficiently combine the detection of particular protein with enormous DNA-based sensing technologies. In this work, the feasibly of the BRCL strategy is first verified through agarose gel electrophoresis and electrochemical impedance spectroscopy measurements, and then confirmed by transferring it to a nanomaterial-assisted fluorescence assay. Significantly, the BRCL strategy-based assay is able to respond to target protein with desirable selectivity, attributing to the specific recognition between small-molecule ligand and its target. Further experiments validate the general applicability of the sensing method by tailoring the ligand toward different proteins (i.e., avidin and folate receptor), and demonstrate its usability in complex biological samples. To our knowledge, this work pioneers the practice of click chemistry in probing specific small-molecule ligand-protein binding, and therefore may pave a new way for selective detection of proteins.
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China; Shanghai Key Laboratory of Bio-Energy Crops, Shanghai University, Shanghai 200444, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Han
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
authors.forename: Zhuxin
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Oncology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. Electronic address: yqshu_njmu@foxmail.com.
authors.forename: Yongqian
authors.initials: Y
authors.lastname: Shu
authors.affiliation: School of Life Sciences, Nanjing University, Nanjing 210093, China. Electronic address: xiangy@nju.edu.cn.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xiang
mesh_terms: D001360:Avidin / Q000737:chemistry; D015374:Biosensing Techniques; D057930:Click Chemistry; D004247:DNA / Q000737:chemistry*; D005453:Fluorescence; D058975:Folate Receptor 1 / Q000737:chemistry; D008024:Ligands; D049329:Nanostructures / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence
chemical_list: D058975:Folate Receptor 1; D008024:Ligands; D011506:Proteins; D001360:Avidin; D004247:DNA
keywords: Click ligation; Clickable DNA probe; Protein detection; Small-molecule ligand