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

Research output: Contribution to journalArticleScientificpeer-review

Researchers

  • Kari Kopra
  • Tanja Seppälä
  • Dana Rabara
  • Maria Abreu-Blanco
  • Sakari Kulmala
  • Matthew Holderfield
  • Harri Härmä

Research units

  • University of Turku
  • Leidos Inc

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.

Details

Original languageEnglish
JournalSensors (Basel, Switzerland)
Volume18
Issue number11
Publication statusPublished - 16 Nov 2018
MoE publication typeA1 Journal article-refereed

    Research areas

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

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