文献类型：期刊文献

英文题名：Tri-Recognition-Mediated Proximity Ligation for Quantitative Analysis of Exosomal Protein-Specific Sialylation and Application on a Microfluidic Platform

作者：Lu, Hongfeng[1];Gu, Zhen[2];Zou, Yilin[1];Wang, Hua[3];Lu, Siyu[1];Wang, Huifeng[2];Ye, Bang-Ce[1];Xu, Huiying[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biosyst & Microanal, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Key Lab Smart Mfg Energy Chem Proc, Minist Educ, Shanghai 200237, Peoples R China;[3]Shanghai Jiao Tong Univ, Renji Hosp, Sch Med, Dept Lab Med, Shanghai 200127, Peoples R China

年份：2025

卷号：97

期号：22

起止页码：11883

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20252218536860);WOS:【SCI-EXPANDED(收录号:WOS:001500120800001)】；

基金：This study was supported by grants from the National Natural Science Foundation of China (22374048, 22134003, and 82472374), National Key Research and Development Program of China (2020YFA0908800), and Natural Science Foundation of Shanghai (22ZR1417800).

语种：英文

外文关键词：Biomarkers - Cell membranes - Signal transduction

摘要：Overexpression of sialylated glycoprotein is a stage-specific process and is regarded as a common manifestation of tumor progression. Accurate quantification of protein-specific sialylation on biological membranes contributes to a thorough comprehension of cellular signal transduction as well as the search for sialylated glycan-related biomarkers. Herein, we propose triple recognition-mediated proximity ligation coupled with rolling-circle amplification to examine protein-specific sialylation on living cell membranes and their derived exosomes. Multiple recognitions in spatial proximity provide three key advantages: (1) significantly improved identification precision, (2) flexible and scalable target options, and (3) minimized off-target effects. Using this approach, we successfully visualize sialylation-dependent interactions between exosomes and cells. By converting certain recognition sites and combining duplex calculations, we establish a quantitation method of exosomal protein-specific sialylation capping ratio, which could act as a useful noninvasive indicator in a customizable 3D-printed microfluidic chip (ExoTRAP) for exosome-based cancer discrimination. This platform enables multiplexed profiling of protein-specific sialylation with high sensitivity (e.g., the LOD of sialylated MUC1-positive MCF-7 exosomes is 2.81 x 106 particles/mL), thus providing new insights into the role of sialylated glycoproteins in exosome functions, as well as a promising strategy for clinical diagnosis.