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Proteomic analyses using an accurate mass and time tag strategy.

Ljiljana Pasa-Tolić1, Christophe Masselon, Richard C Barry

  • 1Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, USA. ljiljana.pasatolic@pnl.gov

Biotechniques
|November 3, 2004
PubMed
Summary

The accurate mass and time (AMT) tag approach enables high-throughput proteomic analysis by creating a validated peptide database. This method enhances efficiency and sensitivity for complex biological samples.

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Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • High-throughput proteomic measurements are crucial for understanding complex biological systems.
  • Existing methods often require extensive validation, limiting throughput.
  • Accurate mass and time (AMT) tagging offers a promising alternative.

Purpose of the Study:

  • To develop and validate the accurate mass and time (AMT) tag approach for high-throughput proteomic analyses.
  • To establish a robust method for creating AMT tag databases.
  • To demonstrate the efficiency and sensitivity of the AMT tag approach.

Main Methods:

  • Creation of an AMT tag database using standard shotgun proteomic analysis.
  • Validation of peptide identifications via Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) and liquid chromatography (LC) elution time.

Related Experiment Videos

  • Utilizing highly reproducible one-dimensional reversed-phased LC separations.
  • Employing FTICR MS for highly accurate mass measurements.

    • Main Results:

    • The AMT tag database approach significantly reduces the need for subsequent MS/MS analyses, enabling high-throughput measurements.
    • Recent improvements allow analysis of picogram amounts of proteome samples with minimized handling and maximized peptide recovery.
    • The nanoproteomics platform can detect over 10^6 differences in protein abundances and identify abundant proteins from subpicogram samples.

    Conclusions:

    • The AMT tag approach is a powerful tool for in-depth and high-throughput proteomic analysis of complex organisms and clinical samples.
    • This technique has the potential to advance proteomic analysis down to the single mammalian cell level.
    • AMT tagging is poised to become a new standard in proteomic research.