Isolation and Quantification of Methylated Cell-Free DNA in Plasma on an Integrated Microfluidic System

Yu Jen Cheng, Chih Hung Wang, Keng Fu Hsu, Gwo Bin Lee

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Methylated cell-free DNA (cfDNA) has been deemed a promising biomarker for ovarian cancer (OvCa) prognosis and therapy selection. However, exploring the methylation profiles of tumor suppressor genes in cfDNA remains a challenge due to their extremely low concentrations and complicated protocols, as well as methodological constraints. In this study, an integrated microfluidic system was developed to automatically (1) capture methylated cfDNA in plasma by magnetic beads coated with the methyl-CpG-binding domain and (2) quantify the methylation level of tumor suppressor genes by on-chip quantitative polymerase chain reaction (qPCR). For capturing methylated cfDNA from a very small amount of plasma, samples along with beads were mixed in a new micromixer to enhance the capture rate. With a high capture rate (72%) and a limit of quantification of 0.1 pg/μL (3 orders of magnitude lower than that of the benchtop method), the compact system could detect the methylated cfDNA from only 20 μL of plasma sample in 2 h. Furthermore, the dynamic range, from 0.1 to 2000 pg/μL of methylated cfDNA, spans the physiological range in plasma, signifying that this device has great potential for personalized medicine in OvCa.

Original languageEnglish
Pages (from-to)2134-2141
Number of pages8
JournalAnalytical Chemistry
Volume94
Issue number4
DOIs
Publication statusPublished - 2022 Feb 1

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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