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

Furst, Ariel L. and Hill, Michael G. and Barton, Jacqueline K. (2013) DNA-Modified Electrodes Fabricated Using Copper-Free Click Chemistry for Enhanced Protein Detection. Langmuir, 29 (52). pp. 16141-16149. ISSN 0743-7463. PMCID PMC3947573. https://resolver.caltech.edu/CaltechAUTHORS:20131224-071741505

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/la403262vDOIArticle
http://pubs.acs.org/doi/abs/10.1021/la403262vPublisherArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3947573/PubMed CentralArticle
ORCID:
AuthorORCID
Barton, Jacqueline K.0000-0001-9883-1600
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.
Funders:
Funding AgencyGrant Number
Office of Naval Research (ONR)N0014-09-1-1117
NIHGM61077
Issue or Number:52
PubMed Central ID:PMC3947573
Record Number:CaltechAUTHORS:20131224-071741505
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20131224-071741505
Official Citation:DNA-Modified Electrodes Fabricated Using Copper-Free Click Chemistry for Enhanced Protein Detection Ariel L. Furst, Michael G. Hill, and Jacqueline K. Barton Langmuir 2013 29 (52), 16141-16149
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:43155
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:24 Dec 2013 17:41
Last Modified:03 Oct 2019 06:05

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