24. ISOLATION AND RECOVERY OF CELL FREE CIRCULATING DNA BIOMARKERS FROM CANCER PATIENT WHOLE BLOOD SAMPLES

Department: Bioengineering
Faculty Advisor(s): Michael Heller
Award(s): Honorable Mention

Primary Student
Name: Avery R Sonnenberg
Email: asonnenb@ucsd.edu
Phone: 716-485-1412
Grad Year: 2012

Abstract
Cell free circulating (cfc) DNA/RNA and other cellular nanoparticulates represent an important class of biomarkers for the early detection of cancer and many other diseases. We have previously developed a novel dielectrophoretic (DEP) method that allows cell free circulating (cfc) DNA, cellular nanoparticulates, and nanoparticles to be directly isolated and detected from undiluted whole blood. At AC frequencies in the 3-10 kHz range and 10 volts peak-to-peak we have shown the separation of cells and micron-sized particles in 1x PBS buffer (Electrophoresis, v. 29, #9, pp. 1765-1774, May 2008) as well as the separation of high molecular weight (hmw) DNA (J. Biophotonics. 2009, 2, 253-261). With the same microarray, we demonstrated isolation of hmw-DNA and fluorescent nanoparticles in the 40 nm to 200 nm range from whole blood. Using this technique we also found visible quantities of SYBR Green fluorescent-stained cfc-DNA and other nanoparticulates in Chronic Lymphocytic Leukemia (CLL) patient blood samples. Based on this work, we have now constructed prototype devices which were designed to increase collection efficiency as well as recoverability of the isolated material. These new devices use sample volumes as small as 25 uL and greatly increase the ability to perform downstream analysis on the collected material, working toward truly ?seamless? sample to answer diagnostics for early cancer and other disease detection.

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