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Updated: Nov 23, 2025

Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging cPILOT
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Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT).

Albert B Arul1, Renã A S Robinson2

  • 1Department of Chemistry, Vanderbilt University.

Journal of Visualized Experiments : Jove
|January 4, 2021
PubMed
Summary

We automated a high-throughput quantitative proteomics workflow (cPILOT) for analyzing up to 24 samples. Automation reduces errors and saves time, making complex proteomics accessible to more researchers.

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Area of Science:

  • Proteomics
  • Mass Spectrometry
  • Biotechnology

Background:

  • Quantitative proteomics enables large-scale protein analysis.
  • Current methods like cPILOT offer high multiplexing but involve manual steps.
  • Manual sample handling can lead to errors, time loss, and reduced reproducibility.

Purpose of the Study:

  • To automate the combined precursor isotopic labeling and isobaric tagging (cPILOT) workflow.
  • To enhance the throughput, reproducibility, and accessibility of quantitative proteomics.
  • To overcome limitations of manual sample processing in high-throughput experiments.

Main Methods:

  • Development and implementation of an automated liquid handling protocol for cPILOT.
  • Utilizing isobaric tagging reagents for multiplexed sample analysis.
  • High-throughput sample preparation for mass spectrometry.

Main Results:

  • Successful transfer of the manual cPILOT protocol to an automated platform.
  • Ability to prepare large sample numbers (96 samples) in parallel.
  • Demonstrated increased feasibility and reproducibility of the cPILOT workflow through automation.

Conclusions:

  • Automation significantly improves the efficiency and reliability of the cPILOT quantitative proteomics workflow.
  • Automated cPILOT enables broader adoption by researchers with automation capabilities.
  • This advancement facilitates more accessible and robust large-scale proteomic studies.