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Improved ATP methodology for biomass assays.

E Schram1, A Weyens-van Witzenburg

  • 1Brussels University, Institute of Molecular Biology, Sint-Genesius-Rode, Belgium.

Journal of Bioluminescence and Chemiluminescence
|July 1, 1989
PubMed
Summary

Standardizing adenosine triphosphate (ATP) methodology improves biomass assays. New methods enhance bacterial ATP extraction and luminometer calibration for greater accuracy and sensitivity in ATP assays.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Microbiology

Background:

  • Adenosine triphosphate (ATP) assays are valuable for biomass assessment but require standardization.
  • Existing ATP methodology faces challenges that can hinder accurate application in biomass assays.
  • Bacteriological applications of ATP assays necessitate specific improvements for reliable results.

Purpose of the Study:

  • To standardize and improve adenosine triphosphate (ATP) methodology for biomass assays.
  • To address limitations in current ATP assay protocols, particularly for bacteriological applications.
  • To propose new methods for enhancing the accuracy, reliability, and sensitivity of ATP measurements.

Main Methods:

  • Replaced apyrase with mammalian ATPase for more specific inhibition during free and somatic ATP destruction.
  • Incorporated lipids to protect luciferase from cationic detergents, ensuring a constant light response during bacterial ATP extraction.
  • Developed new calibration methods for luminometers and sample holder matching in multichannel instruments.

Main Results:

  • Proposed methods facilitate more controlled destruction of free and somatic ATP.
  • Enhanced bacterial ATP extraction yields a constant light response, improving assay consistency.
  • New calibration and matching techniques are presented for luminometer and multichannel instruments.

Conclusions:

  • The proposed modifications significantly improve the standardization and application of ATP methodology in biomass assays.
  • Enhanced methods increase the reliability and sensitivity of ATP assays, particularly in bacteriological contexts.
  • Further development in reagents and instrumentation continues to push the limits of ATP assay sensitivity.

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