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dc.contributor.authorZhou, Wenhu
dc.contributor.authorChen, Qingyun
dc.contributor.authorHuang, Po-Jung Jimmy
dc.contributor.authorDing, Jinsong
dc.contributor.authorLiu, Juewen 20:44:10 (GMT) 20:44:10 (GMT)
dc.descriptionThis document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by publisher. To access the final edited and published work see
dc.description.abstractRNA-cleaving DNAzymes provide a unique platform for developing biosensors. However, a majority of the work has been performed in clean buffer solutions, while the activity of some important DNAzymes in biological sample matrices is still under debate. Two RNA-cleaving DNAzymes (17E and 10-23) are the most widely used. In this work, we carefully studied a few key aspects of the 17E DNAzyme in human blood serum, including hybridization, cleavage activity, and degradation kinetics. Since direct fluorescence monitoring is difficult due to the opacity of serum, denaturing and nondenaturing gel electrophoresis were combined for studying the interaction between serum proteins and DNAzymes. The 17E DNAzyme retains its activity in 90% human blood serum with a cleavage rate of 0.04 min–1, which is similar to that in the PBS buffer (0.06 min–1) with a similar ionic strength. The activity in serum can be accelerated to 0.3 min–1 with an additional 10 mM Ca2+. As compared to 17E, the 10-23 DNAzyme produces negligible cleavage in serum. Degradation of both the substrate and the DNAzyme strand is very slow in serum, especially at room temperature. Degradation occurs mainly at the fluorophore label (linked to DNA via an amide bond) instead of the DNA phosphodiester bonds. Serum proteins can bind more tightly to the 17E DNAzyme complex than to the single-stranded substrate or enzyme. The 17E DNAzyme hybridizes extremely fast in serum. With this understanding, the detection of DNA using the 17E DNAzyme is demonstrated in serum.en
dc.description.sponsorshipUniversity of Waterloo || Natural Sciences and Engineering Research Council || Foundation for Shenghua Scholar of Central South University|| National Natural Science Foundation of China || Grant No. 21301195 Fellowship from the China Scholarship Council || CSC, Grant No. 201406370116en
dc.publisherAmerican Chemical Societyen
dc.subjectDNAzyme Hybridizationen
dc.subjectHuman Blood Serumen
dc.titleDNAzyme Hybridization, Cleavage, Degradation and Sensing in Undiluted Human Blood Serumen
dcterms.bibliographicCitationZhou, W., Chen, Q., Huang, P.-J. J., Ding, J., & Liu, J. (2015). DNAzyme Hybridization, Cleavage, Degradation, and Sensing in Undiluted Human Blood Serum. Analytical Chemistry, 87(7), 4001–4007.
uws.contributor.affiliation1Faculty of Scienceen
uws.contributor.affiliation2Waterloo Institute for Nanotechnology (WIN)en

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