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Reagentless Real-Time ATP Monitoring with New DNA Aptamers.

Adara Bacon1, Obtin Alkhamis1, Caleb Byrd1

  • 1Department of Chemistry, North Carolina State University, 2620 Yarbrough Dr., Raleigh, NC, 27695, USA.

Small (Weinheim an Der Bergstrasse, Germany)
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Summary
This summary is machine-generated.

Researchers developed a new DNA aptamer, ATP18-13, for highly specific and sensitive real-time detection of adenosine triphosphate (ATP). This breakthrough improves upon existing biosensors, enabling precise monitoring in biological samples.

Keywords:
ATPSELEXaptamersmonitoringsensors

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Adenosine triphosphate (ATP) is a critical molecule in biological processes.
  • Existing ATP sensors face limitations in specificity, reagent requirements, and complexity.
  • Current aptamers for ATP detection exhibit insufficient affinity and specificity for practical applications.

Purpose of the Study:

  • To discover novel DNA aptamers with high affinity and specificity for adenosine triphosphate (ATP).
  • To develop an improved aptamer-based biosensor for real-time ATP detection.
  • To demonstrate the sensor's utility in monitoring biological processes involving ATP.

Main Methods:

  • Systematic evolution of ligands by exponential enrichment (SELEX) was employed to identify new aptamers.
  • The best-performing aptamer, ATP18-13, was selected based on affinity and specificity.
  • A fluorescent beacon sensor was constructed using ATP18-13 for ATP detection.

Main Results:

  • The novel aptamer ATP18-13 exhibits significantly higher affinity for ATP compared to existing aptamers.
  • ATP18-13 demonstrates superior specificity, effectively discriminating against adenosine diphosphate (ADP), adenosine monophosphate (AMP), and adenosine.
  • The developed fluorescent beacon sensor achieved a limit of detection of 125 nm ATP in buffer and 1 µm in cell culture medium.
  • Real-time monitoring of deoxyATP consumption during DNA polymerase activity was successfully demonstrated.

Conclusions:

  • The newly discovered DNA aptamers offer unprecedented affinity and specificity for ATP detection.
  • The developed aptamer-based sensor enables sensitive and real-time monitoring of ATP.
  • These aptamers and sensors hold promise for diverse biological applications requiring ATP measurement.