24 August 2010 Rapid detection of cancer related DNA nanoparticulate biomarkers and nanoparticles in whole blood
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Abstract
The ability to rapidly detect cell free circulating (cfc) DNA, cfc-RNA, exosomes and other nanoparticulate disease biomarkers as well as drug delivery nanoparticles directly in blood is a major challenge for nanomedicine. We now show that microarray and new high voltage dielectrophoretic (DEP) devices can be used to rapidly isolate and detect cfc-DNA nanoparticulates and nanoparticles directly from whole blood and other high conductance samples (plasma, serum, urine, etc.). At DEP frequencies of 5kHz-10kHz both fluorescent-stained high molecular weight (hmw) DNA, cfc-DNA and fluorescent nanoparticles separate from the blood and become highly concentrated at specific DEP highfield regions over the microelectrodes, while blood cells move to the DEP low field-regions. The blood cells can then be removed by a simple fluidic wash while the DNA and nanoparticles remain highly concentrated. The hmw-DNA could be detected at a level of <260ng/ml and the nanoparticles at <9.5 x 109 particles/ml, detection levels that are well within the range for viable clinical diagnostics and drug nanoparticle monitoring. Disease specific cfc-DNA materials could also be detected directly in blood from patients with Chronic Lymphocytic Leukemia (CLL) and confirmed by PCR genotyping analysis.
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Michael J. Heller, Michael J. Heller, Raj Krishnan, Raj Krishnan, Avery Sonnenberg, Avery Sonnenberg, "Rapid detection of cancer related DNA nanoparticulate biomarkers and nanoparticles in whole blood", Proc. SPIE 7759, Biosensing III, 77590P (24 August 2010); doi: 10.1117/12.861579; https://doi.org/10.1117/12.861579
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