Dielectrophoretic isolation of DNA and nanoparticles from blood

被引:60
作者
Sonnenberg, Avery [2 ]
Marciniak, Jennifer Y. [3 ]
Krishnan, Rajaram [4 ]
Heller, Michael J. [1 ,2 ]
机构
[1] Univ Calif San Diego, Dept Nanoengn, La Jolla, CA 92093 USA
[2] Univ Calif San Diego, Dept Bioengn, La Jolla, CA 92093 USA
[3] UCSD Moores Canc Ctr, La Jolla, CA USA
[4] Biol Dynam, La Jolla, CA USA
关键词
Biomarkers; CFC-DNA; Dielectrophoresis; High conductance; Nanoparticles; Whole blood; CIRCULATING TUMOR DNA; AC ELECTROKINETICS; CANCER-PATIENTS; NUCLEIC-ACIDS; CELLS; SEPARATION; PLASMA; PARTICLES; QUANTIFICATION; NANOCARRIERS;
D O I
10.1002/elps.201100700
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 x 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.
引用
收藏
页码:2482 / 2490
页数:9
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