Ligand Dilution Analysis Facilitates Aptamer Binding Characterization at the Single‐Molecule Level. Issue 10 (29th December 2022)

• Record Type: Journal Article
• Authors: Du, Yulin; Lyu, Yifan; Li, Shiquan; Ding, Ding; Chen, Jianghuai; Yang, Cai; Sun, Yang; Qu, Fengli; Xiao, Zeyu; Jiang, Jianhui; Tan, Weihong
• Is Part Of: Angewandte Chemie international edition. Volume 62:Issue 10(2023)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-12-29
• Subjects: Aptamers -- Binding Site -- Dilution -- Fluorescent Probes -- Motion Analysis; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.202215387
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Cell‐specific aptamers offer a powerful tool to study membrane receptors at the single‐molecule level. Most target receptors of aptamers are highly expressed on the cell surface, but difficult to analyze in situ because of dense distribution and fast velocity. Therefore, we herein propose a random sampling‐based analysis strategy termed ligand dilution analysis (LDA) for easily implemented aptamer‐based receptor study. Receptor density on the cell surface can be calculated based on a regression model. By using a synergistic ligand dilution design, colocalization and differentiation of aptamer and monoclonal antibody (mAb) binding on a single receptor can be realized. Once this is accomplished, precise binding site and detailed aptamer‐receptor binding mode can be further determined using molecular docking and molecular dynamics simulation. The ligand dilution strategy also sets the stage for an aptamer‐based dynamics analysis of two‐ and three‐dimensional motion and fluctuation of highly expressed receptors on the live cell membrane. Abstract : An easily implemented membrane‐receptor‐analysis method termed ligand dilution analysis, combining the concepts of single‐molecule localization and random sampling, is reported. Aptamer binding characterization, including receptor density calculation, single‐molecule colocalization, binding site determination, motion and fluctuation analysis, is performed with minimal probe and instrument requirements.