Paper/PMMA hybrid device with a microvalve-controlled design for exosome isolation and analysis

Wen-Pin Hu; Yi-Fang Lai; Cao-An Vu; Chia-Wen Tsao; Shin-Chen Pan; Chao-Min Cheng; Wen-Yih Chen

doi:10.1016/j.talanta.2023.124851

Paper/PMMA hybrid device with a microvalve-controlled design for exosome isolation and analysis

Talanta. 2023 Dec 1:265:124851. doi: 10.1016/j.talanta.2023.124851. Epub 2023 Jun 20.

Authors

Wen-Pin Hu¹, Yi-Fang Lai², Cao-An Vu², Chia-Wen Tsao³, Shin-Chen Pan⁴, Chao-Min Cheng⁵, Wen-Yih Chen⁶

Affiliations

¹ Department of Bioinformatics and Medical Engineering, Asia University, Taichung, 41354, Taiwan; Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, 41354, Taiwan.
² Department of Chemical and Materials Engineering, National Central University, Jhong-Li, 32001, Taiwan.
³ Department of Mechanical Engineering, National Central University, Jhong-Li, 32001, Taiwan.
⁴ Department of Surgery, Section of Plastic and Reconstructive Surgery, Center of Cell Therapy, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, 704, Taiwan.
⁵ Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
⁶ Department of Chemical and Materials Engineering, National Central University, Jhong-Li, 32001, Taiwan. Electronic address: wychen@ncu.edu.tw.

PMID: 37354627
DOI: 10.1016/j.talanta.2023.124851

Abstract

This study proposes a paper/PMMA hybrid device designed to isolate exosomes and extract exosomal miRNA, followed by quantitative analysis. It aims to provide simplified and convenient sample preparation for potential point-of-care testing (POCT) processes. In contrast to previous work conducted by our research team, which focused on isolating exosomes and exosomal nucleic acids, this study introduces a novel approach by integrating paper and a PMMA mold with a microvalve controlled design. This innovative method enables the entire process to be performed on paper. The pressure on the paper could be adjusted by turning the screw upon the valve to change the pore size and permeability of the paper, which achieved the effect of controlling the flow rate of fluids. The paper was designed to have an immunoaffinity area for capturing exosomes and a sol-gel silica coating area for extracting miRNA. The paper-based ELISA (p-ELISA) exhibited a limit of detection and a limit of quantitation of 6 × 10⁷ and 5.4 × 10⁸ particles/mL, respectively, for exosome measurement. The reverse transcription quantitative polymerase chain reaction (RT-qPCR) revealed that the Ct (threshold cycle) value for quantifying the miR-21 in the miRNAs extracted by the proposed paper/PMMA hybrid device was comparable to the Ct value of the commercial extraction kit. The developed paper/PMMA hybrid device with a microvalve-controlled design should be incorporated into the POCT system to extract exosomal miRNAs.

Keywords: Exosomal miRNA; Microvalve-controlled design; Paper/PMMA hybrid device; miRNA extraction; microRNA-21 (miR-21).

MeSH terms

Exosomes* / chemistry
MicroRNAs* / analysis
Polymethyl Methacrylate

Substances

Polymethyl Methacrylate
MicroRNAs