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Volume 7, Issue 6(Suppl) J Chromatogr Sep Tech
ISSN: 2157-7064 JCGST, an open access journal
Page 30
Notes:
Separation Techniques 2016
September 26-28, 2016
conferenceseries.com
Separation Techniques
September 26-28, 2016 Valencia, Spain
2nd International Conference and Expo on
Identification of inhibitors of the Fe-2OG dependent oxygenases by capillary electrophoresis with
laser-induced fluorescence
S M Krylova
York University, Canada
2-oxoglutarate and Fe (II)-dependent dioxygenases catalyze the removal of N-alkyl groups from damaged DNA in eukaryotes
and bacteria. Silencing these demethylating enzymes may be beneficial for the enhancement of chemotherapeutic treatments
and reduction of their cytotoxic effects. Therefore, a direct and efficient quantitative analysis using biologically-relevant
substrates is needed for detection of demethylase activity of E. coli and human Fe-2OG dioxygenases and its application for
high throughput screening of potential inhibitors. Previously reported techniques utilize coupled enzyme reactions, detect co-
products, require complex processing or use radioactive substances. We developed a direct and rapid method based on capillary
electrophoresis with laser-induced detection allowing real-time detection of both the substrate and the product separated with
high efficiency using no post-enzymatic processing and without the use of radioactive substances. Here we report the CE-based
activity assay of two members of the Fe-2OG dioxygenase superfamily of enzymes-hABH2 and hABH3, and demonstrate that
the activity can be selectively inhibited by small molecules or short DNA aptamers. The inhibition selectivity of hABH2 over
hABH3 enzymes can be advantageously used for qualitative and quantitative assay of these enzymes mixtures. The simple and
specific differential analysis can be potentially employed to distinguish hABH2 and hABH3 enzymes expressed by the same
types of cells in vivo. The minimal processing, short analysis time, low cost and availability of automation makes this assay
useful for developing therapies targeting Fe-2OG dioxygenases.
Biography
S M Krylova has obtained her PhD from the Russian Academy of Sciences, Russia. She has over 10 years of research leadership experience in the area of Medical
Diagnostics and Drug Development in biotechnology and pharmaceutical companies in Canada. She has been a contract Faculty Member at York University in
Toronto since 2008. She is also leading research projects in the area of Bioanalytical Chemistry as a Senior Research Associate in the Centre for Research on
Biomolecular Interactions at York University.
S M Krylova, J Chromatogr Sep Tech 2016, 7:6(Suppl)