Enzymatically amplified SPR imaging detection of DNA by exonuclease III digestion of DNA microarrays

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Lee, H.J. and Li, Y. and Wark, Alastair and Corn, R.M. (2005) Enzymatically amplified SPR imaging detection of DNA by exonuclease III digestion of DNA microarrays. Analytical Chemistry, 77 (16). pp. 5096-5100. ISSN 0003-2700 (https://doi.org/10.1021/ac050815w)

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Abstract

This paper describes a novel approach utilizing the enzyme exonuclease III in conjunction with 3‘-terminated DNA microarrays for the amplified detection of single-stranded DNA (ssDNA) with surface plasmon resonance (SPR) imaging. When ExoIII and target DNA are simultaneously introduced to a 3‘-terminated ssDNA microarray, hybridization adsorption of the target ssDNA leads to the direction-dependent ExoIII hydrolysis of probe ssDNA strands and the release of the intact target ssDNA back into the solution. Readsorption of the target ssDNA to another probe creates a repeated hydrolysis process that results over time in a significant negative change in SPR imaging signal. Experiments are presented that demonstrate the direction-dependent surface enzyme reaction of ExoIII with double-stranded DNA as well as this new enzymatically amplified SPR imaging process with a 16-mer target ssDNA detection limit of 10−100 pM. This is a 102−103 improvement on previously reported measurements of SPR imaging detection of ssDNA based solely on hybridization adsorption without enzymatic amplification

ORCID iDs

Lee, H.J., Li, Y., Wark, Alastair ORCID logoORCID: https://orcid.org/0000-0001-8736-7566 and Corn, R.M.;
CORE (COnnecting REpositories)