Frontiers in Bioengineering and Biotechnology | |
Paper-Based Devices for Capturing Exosomes and Exosomal Nucleic Acids From Biological Samples | |
Chih-Ling Lee1  Wen-Yih Chen1  Chi-Hung Lai1  Cao-An Vu1  Van-Truc Vu1  Chao-Min Cheng2  Yao-Hung Tsai2  | |
[1] Department of Chemical and Materials Engineering, National Central University, Taoyuan, Taiwan;Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan; | |
关键词: paper-based device; exosome; exosomal miRNA; nucleic acid extraction; immunoassay; plasma; | |
DOI : 10.3389/fbioe.2022.836082 | |
来源: DOAJ |
【 摘 要 】
Exosomes, nanovesicles derived from cells, contain a variety of biomolecules that can be considered biomarkers for disease diagnosis, including microRNAs (miRNAs). Given knowledge and demand, inexpensive, robust, and easy-to-use tools that are compatible with downstream nucleic acid detection should be developed to replace traditional methodologies for point-of-care testing (POCT) applications. This study deploys a paper-based extraction kit for exosome and exosomal miRNA analytical system with some quantifying methods to serve as an easy sample preparation for a possible POCT process. Exosomes concentrated from HCT116 cell cultures were arrested on paper-based immunoaffinity devices, which were produced by immobilizing anti-CD63 antibodies on Whatman filter paper, before being subjected to paper-based silica devices for nucleic acids to be trapped by silica nanoparticles adsorbed onto Whatman filter paper. Concentrations of captured exosomes were quantified by enzyme-linked immunosorbent assay (ELISA), demonstrating that paper-based immunoaffinity devices succeeded in capturing and determining exosome levels from cells cultured in both neutral and acidic microenvironments, whereas microRNA 21 (miR-21), a biomarker for various types of cancers and among the nucleic acids absorbed onto the silica devices, was determined by reverse transcription quantitative polymerase chain reaction (RT-qPCR) to prove that paper-based silica devices were capable of trapping exosomal nucleic acids. The developed paper-based kit and the devised procedure was successfully exploited to isolate exosomes and exosomal nucleic acids from different biological samples (platelet-poor plasma and lesion fluid) as clinical applications.
【 授权许可】
Unknown