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Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content

1
Materials Chemistry and Chemical Analysis, University of Turku, Vatselankatu 2, 20500 Turku, Finland
2
NCI-RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD 21702, USA
3
Laboratory of Analytical Chemistry, Department of Chemistry, Aalto University, P.O. Box 16100, 00076 Aalto, Finland
*
Author to whom correspondence should be addressed.
Sensors 2018, 18(11), 3989; https://doi.org/10.3390/s18113989
Received: 3 October 2018 / Revised: 11 November 2018 / Accepted: 12 November 2018 / Published: 16 November 2018
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Abstract

A new label-free molecular probe for luminescent nucleotide detection in neutral aqueous solution is presented. Phosphate-containing molecules, such as nucleotides possess vital role in cell metabolism, energy economy, and various signaling processes. Thus, the monitoring of nucleotide concentration and nucleotide related enzymatic reactions is of high importance. Two component lanthanide complex formed from Tb(III) ion carrier and light harvesting antenna, readily distinguishes nucleotides containing different number of phosphates and enable direct detection of enzymatic reactions converting nucleotide triphosphate (NTP) to nucleotide di/monophosphate or the opposite. Developed sensor enables the detection of enzymatic activity with a low nanomolar sensitivity, as highlighted with K-Ras and apyrase enzymes in their hydrolysis assays performed in a high throughput screening compatible 384-well plate format. View Full-Text
Keywords: nucleotide triphosphate; label-free; K-Ras; apyrase; terbium nucleotide triphosphate; label-free; K-Ras; apyrase; terbium
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Kopra, K.; Seppälä, T.; Rabara, D.; Abreu-Blanco, M.; Kulmala, S.; Holderfield, M.; Härmä, H. Label-Free Time-Gated Luminescent Detection Method for the Nucleotides with Varying Phosphate Content. Sensors 2018, 18, 3989.

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