Dielectrophoretic isolation and detection of cancer-related circulating cell-free DNA biomarkers from blood and plasma

Avery Sonnenberg, Jennifer Y. Marciniak, Elaine A. Skowronski, Sareh Manouchehri, Laura Rassenti, Emanuela M. Ghia, George F. Widhopf, Thomas J. Kipps, Michael J. Heller

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

Conventional methods for the isolation of cancer-related circulating cell-free (ccf) DNA from patient blood (plasma) are time consuming and laborious. A DEP approach utilizing a microarray device now allows rapid isolation of ccf-DNA directly from a small volume of unprocessed blood. In this study, the DEP device is used to compare the ccf-DNA isolated directly from whole blood and plasma from 11 chronic lymphocytic leukemia (CLL) patients and one normal individual. Ccf-DNA from both blood and plasma samples was separated into DEP high-field regions, after which cells (blood), proteins, and other biomolecules were removed by a fluidic wash. The concentrated ccf-DNA was detected on-chip by fluorescence, and then eluted for PCR and DNA sequencing. The complete process from blood to PCR required less than 10 min; an additional 15 min was required to obtain plasma from whole blood. Ccf-DNA from the equivalent of 5 μL of CLL blood and 5 μL of plasma was amplified by PCR using Ig heavy-chain variable (IGHV) specific primers to identify the unique IGHV gene expressed by the leukemic B-cell clone. The PCR and DNA sequencing results obtained by DEP from all 11 CLL blood samples and from 8 of the 11 CLL plasma samples were exactly comparable to the DNA sequencing results obtained from genomic DNA isolated from CLL patient leukemic B cells (gold standard).

Original languageEnglish (US)
Pages (from-to)1828-1836
Number of pages9
JournalELECTROPHORESIS
Volume35
Issue number12-13
DOIs
StatePublished - Jul 2014

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Keywords

  • Biomarkers
  • Cancer
  • Chronic lymphocytic leukemia (CLL)
  • Circulating cell-free (ccf)-DNA
  • Dielectrophoresis

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

Cite this

Sonnenberg, A., Marciniak, J. Y., Skowronski, E. A., Manouchehri, S., Rassenti, L., Ghia, E. M., Widhopf, G. F., Kipps, T. J., & Heller, M. J. (2014). Dielectrophoretic isolation and detection of cancer-related circulating cell-free DNA biomarkers from blood and plasma. ELECTROPHORESIS, 35(12-13), 1828-1836. https://doi.org/10.1002/elps.201400016