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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 FROM AUTHOR]

Aptamer binding characterization, including receptor density calculation, single-molecule colocalization, binding site determination, motion and fluctuation analysis, was performed with minimal probe and instrument requirements. Aptamers, Binding Site, Dilution, Fluorescent Probes, Motion Analysis Keywords: Aptamers; Binding Site; Dilution; Fluorescent Probes; Motion Analysis EN Aptamers Binding Site Dilution Fluorescent Probes Motion Analysis 1 1 1 02/22/23 20230301 NES 230301 B A membrane-receptor-analysis method b termed ligand dilution analysis, which combines the concepts of single-molecule localization and random sampling, is reported by Yifan Lyu, Jianhui Jiang, Weihong Tan et al. in their Research Article (e202215387). [Extracted from the article]

Aptamer binding characterization, including receptor density calculation, single-molecule colocalization, binding site determination, motion and fluctuation analysis, was performed with minimal probe and instrument requirements. Back Cover: Ligand Dilution Analysis Facilitates Aptamer Binding Characterization at the Single-Molecule Level (Angew. Keywords: Aptamers; Binding Site; Dilution; Fluorescent Probes; Motion Analysis EN Aptamers Binding Site Dilution Fluorescent Probes Motion Analysis 1 1 1 02/22/23 20230301 NES 230301 B A membrane-receptor-analysis method b termed ligand dilution analysis, which combines the concepts of single-molecule localization and random sampling, is reported by Yifan Lyu, Jianhui Jiang, Weihong Tan et al. in their Research Article (e202215387). [Extracted from the article]