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Published December 31, 2013 | Supplemental Material + Accepted Version
Journal Article Open

DNA-Modified Electrodes Fabricated Using Copper-Free Click Chemistry for Enhanced Protein Detection

Abstract

A method of DNA monolayer formation has been developed using copper-free click chemistry that yields enhanced surface homogeneity and enables variation in the amount of DNA assembled; extremely low-density DNA monolayers, with as little as 5% of the monolayer being DNA, have been formed. These DNA-modified electrodes (DMEs) were characterized visually, with AFM, and electrochemically, and were found to facilitate DNA-mediated reduction of a distally bound redox probe. These low-density monolayers were found to be more homogeneous than traditional thiol-modified DNA monolayers, with greater helix accessibility through an increased surface area-to-volume ratio. Protein binding efficiency of the transcriptional activator TATA-binding protein (TBP) was also investigated on these surfaces and compared to that on DNA monolayers formed with standard thiol-modified DNA. Our low-density monolayers were found to be extremely sensitive to TBP binding, with a signal decrease in excess of 75% for 150 nM protein. This protein was detectable at 4 nM, on the order of its dissociation constant, with our low-density monolayers. The improved DNA helix accessibility and sensitivity of our low-density DNA monolayers to TBP binding reflects the general utility of this method of DNA monolayer formation for DNA-based electrochemical sensor development.

Additional Information

© 2013 American Chemical Society. Received: September 3, 2013; Revised: October 30, 2013. Publication Date (Web): December 11, 2013. We are grateful to the Office of Naval Research (N0014-09-1-1117) and the NIH (GM61077) for their financial support. We also thank Prof. Eileen Spain of Occidental College for input and helpful discussion for the AFM studies.

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Accepted Version - nihms548814.pdf

Supplemental Material - la403262v_si_001.pdf

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