Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

Kari Kopra
* ORCID logo ,
Tanja Seppälä
,
Dana Rabara
,
Maria Abreu-Blanco
,
Sakari Kulmala
ORCID logo ,
Matthew Holderfield
,
Harri Härmä
Version 1 : Received: 3 October 2018 / Approved: 3 October 2018 / Online: 3 October 2018 (17:32:44 CEST)

A peer-reviewed article of this Preprint also exists.

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. 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.

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 signaling. 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 there hydrolysis assays performed in high throughput screening compatible 384-well plate format.

Keywords

nucleotide triphosphate, label-free, K-Ras, apyrase, terbium

Subject

Biology and Life Sciences, Biology and Biotechnology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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