Evolution of Cross-Linking Aptamers for Long-Lasting Membrane Protein Labeling
Siqi Bian, Qibing Qiu, Chenshuo Zhang, Haiyan Cao, Hong Xuan, Xueling Yan, Jun Li, Wenlong Chen, Xiyang Liu, Yifan Chen, Jinchen Long, Huang Su, Jing Wang, Liqin Zhang
Abstract
The growing demand for membrane protein research, coupled with the structural flexibility and high-throughput screening efficiency of aptamers as ligand candidates, has established aptamers as a robust tool for investigating membrane proteins. In this study, we present a novel in vitro evolution strategy for the development of covalent aptamers, leveraging the principles of fluorine-sulfur exchange (SuFEx) chemistry. This strategy facilitates the screening of aptamers with high specificity for target membrane proteins, ultimately yielding cross-linking aptamers (xl-aptamers). These aptamers exhibit sustained and specific binding affinity toward their target proteins, thereby enabling efficient cell classification. Their performance underscores considerable potential for applications in single-cell and spatial proteomics research.