Dielectrophoretic isolation of DNA and nanoparticles from blood

The ability to effectively detect disease‐related DNA biomarkers and drug delivery nanoparticles directly in blood is a major challenge for viable diagnostics and therapy monitoring. A DEP method has been developed which allows the rapid isolation, concentration and detection of DNA and nanoparticles directly from human and rat whole blood. Using a microarray device operating at 20 V peak‐to‐peak and 10 kHz, a wide range of high molecular weight (HMW)‐DNA and nanoparticles were concentrated into high‐field regions by positive DEP, while the blood cells were concentrated into the low‐field regions by negative DEP. A simple fluidic wash removes the blood cells while the DNA and nanoparticles remain concentrated in the DEP high‐field regions where they can be detected by fluorescence. HMW‐DNA could be detected at 260 ng/mL, which is a detection level suitable for analysis of disease‐related cell‐free circulating DNA biomarkers. Fluorescent 40 nm nanoparticles could be detected at 9.5 × 109 particles/mL, which is a level suitable for monitoring drug delivery nanoparticles. The ability to rapidly isolate and detect DNA biomarkers and nanoparticles from undiluted whole blood will benefit many diagnostic applications by significantly reducing sample preparation time and complexity.

[1]  Kaela M. Leonard,et al.  Explorations of ABO‐Rh antigen expressions on erythrocyte dielectrophoresis: Changes in cross‐over frequency , 2011, Electrophoresis.

[2]  Hsueh-Chia Chang,et al.  Optimized DNA hybridization detection on nanocolloidal particles by dielectrophoresis , 2010, Electrophoresis.

[3]  G van den Engh,et al.  Trapping of DNA in nonuniform oscillating electric fields. , 1998, Biophysical journal.

[4]  大房 健.  基礎講座 電気泳動(Electrophoresis) , 2005 .

[5]  G. van den Engh,et al.  Trapping of DNA by dielectrophoresis , 2002, Electrophoresis.

[6]  F. O. Fackelmayer,et al.  DNA fragments in the blood plasma of cancer patients: quantitations and evidence for their origin from apoptotic and necrotic cells. , 2001, Cancer research.

[7]  H. Morgan,et al.  Ac electrokinetics: a survey of sub-micrometre particle dynamics , 2000 .

[8]  Mandy L Y Sin,et al.  Hybrid electrokinetic manipulation in high-conductivity media. , 2011, Lab on a chip.

[9]  H Morgan,et al.  Separation of submicron bioparticles by dielectrophoresis. , 1999, Biophysical journal.

[10]  M. Heller,et al.  Rapid determination of single base mismatch mutations in DNA hybrids by direct electric field control. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Robert L Sah,et al.  Probing the role of multicellular organization in three-dimensional microenvironments , 2006, Nature Methods.

[12]  F F Becker,et al.  Separation of human breast cancer cells from blood by differential dielectric affinity. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. Heller,et al.  Preparation and hybridization analysis of DNA/RNA from E. coli on microfabricated bioelectronic chips , 1998, Nature Biotechnology.

[14]  Ion Stiharu,et al.  Interdigitated comb‐like electrodes for continuous separation of malignant cells from blood using dielectrophoresis , 2011, Electrophoresis.

[15]  Darrell J Irvine,et al.  Drug delivery: One nanoparticle, one kill. , 2011, Nature materials.

[16]  Z. Hall.  Cancer , 1906, The Hospital.

[17]  M. Sano,et al.  Contactless dielectrophoretic spectroscopy: Examination of the dielectric properties of cells found in blood , 2011, Electrophoresis.

[18]  Magnus Willander,et al.  Trapping single molecules by dielectrophoresis. , 2005, Physical review letters.

[19]  M. Ferrari.  Cancer nanotechnology: opportunities and challenges , 2005, Nature Reviews Cancer.

[20]  J. Milner,et al.  Dielectrophoretic investigation of plant virus particles: Cow Pea Mosaic Virus and Tobacco Mosaic Virus , 2006, Electrophoresis.

[21]  Sadik C Esener,et al.  Interaction of Nanoparticles at the DEP Microelectrode Interface under High Conductance Conditions. , 2009, Electrochemistry communications.

[22]  Saeid Nahavandi,et al.  Dielectrophoretic platforms for bio-microfluidic systems. , 2011, Biosensors & bioelectronics.

[23]  M J Hawkins,et al.  Abraxane, a novel Cremophor-free, albumin-bound particle form of paclitaxel for the treatment of advanced non-small-cell lung cancer. , 2006, Annals of oncology : official journal of the European Society for Medical Oncology.

[24]  Jason P. Gleghorn,et al.  Rare Cell Capture in Microfluidic Devices. , 2011, Chemical engineering science.

[25]  M. Stroun,et al.  Isolation and characterization of DNA from the plasma of cancer patients. , 1987, European journal of cancer & clinical oncology.

[26]  Sadik C Esener,et al.  Alternating current electrokinetic separation and detection of DNA nanoparticles in high‐conductance solutions , 2008, Electrophoresis.

[27]  Frank Diehl,et al.  Detection and quantification of mutations in the plasma of patients with colorectal tumors. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Caroline Dive,et al.  DNA Methylation in Circulating Tumour DNA as a Biomarker for Cancer , 2007, Biomarker insights.

[29]  U. Pastorino,et al.  Quantification of Free Circulating DNA As a Diagnostic Marker in Lung Cancer , 2003 .

[30]  Nobuhiro Nishiyama,et al.  Nanomedicine: nanocarriers shape up for long life. , 2007, Nature nanotechnology.

[31]  K. R. Visser.  Electric conductivity of stationary and flowing human blood at low frequencies , 2006, Medical and Biological Engineering and Computing.

[32]  Antonio Juárez,et al.  On‐line separation of bacterial cells by carbon‐electrode dielectrophoresis , 2010, Electrophoresis.

[33]  James T. Wu,et al.  Cell-free DNA: measurement in various carcinomas and establishment of normal reference range. , 2002, Clinica chimica acta; international journal of clinical chemistry.

[34]  M. Heller,et al.  Isolation of cultured cervical carcinoma cells mixed with peripheral blood cells on a bioelectronic chip. , 1998, Analytical chemistry.

[35]  Kazunori Kataoka,et al.  Current state, achievements, and future prospects of polymeric micelles as nanocarriers for drug and gene delivery. , 2006, Pharmacology & therapeutics.

[36]  Soumya K. Srivastava,et al.  Direct current insulator‐based dielectrophoretic characterization of erythrocytes: ABO‐Rh human blood typing , 2011, Electrophoresis.

[37]  R. Stahel,et al.  Circulating DNA: a new diagnostic gold mine? , 2002, Cancer treatment reviews.

[38]  A. B. Frazier,et al.  Lateral-driven continuous dielectrophoretic microseparators for blood cells suspended in a highly conductive medium. , 2008, Lab on a chip.

[39]  M. Fleischhacker,et al.  Circulating nucleic acids (CNAs) and cancer--a survey. , 2007, Biochimica et Biophysica Acta.

[40]  H Morgan,et al.  The dielectrophoretic levitation and separation of latex beads in microchips , 2001, Electrophoresis.

[41]  F G HIRSCH,et al.  The electrical conductivity of blood. I: Relationship to erythrocyte concentration. , 1950, Blood.

[42]  Hsueh-Chia Chang,et al.  Dielectrophoretic microfluidic device for the continuous sorting of Escherichia coli from blood cells. , 2011, Biomicrofluidics.

[43]  Walter Weder,et al.  Circulating deoxyribonucleic Acid as prognostic marker in non-small-cell lung cancer patients undergoing chemotherapy. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[44]  P. Vineis,et al.  Circulating free DNA in plasma or serum as biomarker of carcinogenesis: practical aspects and biological significance. , 2007, Mutation research.

[45]  Donald Wlodkowic,et al.  Dynamic analysis of drug-induced cytotoxicity using chip-based dielectrophoretic cell immobilization technology. , 2011, Analytical chemistry.

[46]  L. Mariani,et al.  Analysis of circulating tumor DNA in plasma at diagnosis and during follow-up of lung cancer patients. , 2001, Cancer research.

[47]  R. Pethig,et al.  The dielectrophoresis enrichment of CD34+ cells from peripheral blood stem cell harvests. , 1996, Bone marrow transplantation.

[48]  Klaus Pantel,et al.  Cell-free nucleic acids as biomarkers in cancer patients , 2011, Nature Reviews Cancer.

[49]  J. Jen,et al.  An Overview on the Isolation and Analysis of Circulating Tumor DNA in Plasma and Serum , 2000, Annals of the New York Academy of Sciences.

[50]  Chin-Hong Chen,et al.  Does Conductance Catheter Measurement System Give Consistent and Reliable Pressure–Volume Relations in Rats? , 2011, IEEE Transactions on Biomedical Engineering.

[51]  H. Morgan,et al.  Ac electrokinetics: a review of forces in microelectrode structures , 1998 .

[52]  Richard Superfine,et al.  Two-Dimensional Manipulation and Orientation of Actin−Myosin Systems with Dielectrophoresis , 2003 .

[53]  Michael J Heller,et al.  An AC electrokinetic method for enhanced detection of DNA nanoparticles. , 2009, Journal of biophotonics.

[54]  H. Morgan,et al.  The electrokinetic properties of latex particles: comparison of electrophoresis and dielectrophoresis. , 2005, Journal of colloid and interface science.