Journal
SENSORS AND ACTUATORS A-PHYSICAL
Volume 195, Issue -, Pages 183-190Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.sna.2012.07.022
Keywords
Solid-phase extraction; Electrophoresis; Microchannel; ssDNA; Human immunoglobulin E
Funding
- National Science Foundation [CBET-0854030]
- National Institutes of Health [RR025816-02, CA147925-01]
- Directorate For Engineering
- Div Of Chem, Bioeng, Env, & Transp Sys [0854030] Funding Source: National Science Foundation
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This paper presents a microchip that isolates and enriches target-binding single-stranded DNA (ssDNA) from a randomized DNA mixture using a combination of solid-phase extraction and electrophoresis. Strands of ssDNA in a randomized mixture are captured via specific binding onto target-functionalized microbeads in a microchamber. The strands are further separated from impurities and enriched on-chip via electrophoresis. The microchip consists of two microchambers that are connected by a channel filled with agarose gel. In the isolation chamber, beads functionalized with human immunoglobulin E (IgE) are retained by a weir structure. An integrated heater elevates the temperature in the chamber to elute desired ssDNA from the beads, and electrophoretic transport of the DNA through the gel to the second chamber is accomplished by applying an electric potential difference between the two chambers. Experimental results show that ssDNA expressing binding affinity to IgE was captured and enriched from a sample of ssDNA with random sequences, demonstrating the potential of the microchip to enhance the sensitivity of ssDNA detection methods in dilute and complex biological samples. (C) 2012 Elsevier B.V. All rights reserved.
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