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Updated: Jul 4, 2026

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High-Throughput, Ultralow-Input Proteomics Enabled by Narrow-Bore Open Tubular Liquid Chromatography.

Piliang Xiang1, Yumi Kwon1, Ljiljana Paša-Tolić1

  • 1Environmental Molecular Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99354, United States.

Analytical Chemistry
|July 2, 2026
PubMed
Summary

Researchers developed a new method for picogram-scale proteomics, significantly improving sample injection for narrow bore open tubular liquid chromatography (nOTLC). This breakthrough enhances both speed and sensitivity for analyzing limited biological samples.

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Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biotechnology

Background:

  • Picogram-scale proteomics is crucial for studying cellular heterogeneity but faces challenges in balancing sensitivity and throughput.
  • Liquid chromatography (LC) performance limitations significantly impact the efficiency of low-input proteomics.
  • Existing nOTLC methods suffer from high sample loss during injection, hindering their application.

Purpose of the Study:

  • To address the sensitivity-throughput trade-off in picogram-scale proteomics.
  • To implement a low-loss sample injection method for narrow bore open tubular LC (nOTLC).
  • To optimize an nOTLC-MS system for high-throughput, high-sensitivity proteomic analysis.

Main Methods:

  • Development and implementation of a low-loss sample injection technique for nOTLC.

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Published on: May 20, 2013

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Last Updated: Jul 4, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
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Published on: May 20, 2013

  • Utilizing narrow bore open tubular LC coupled with mass spectrometry (nOTLC-MS).
  • Analysis of HeLa digest samples at picogram levels (<40 pg).

    • Main Results:

    • Achieved a 295-fold improvement in sample injection efficiency compared to previous nOTLC methods.
    • Identified an average of 3955 proteins from <40 pg of HeLa digest at 261 samples per day (SPD).
    • Demonstrated potential for 720 SPD with identification of 2460 proteins from ~20 pg of HeLa digest, showcasing enhanced sensitivity and throughput.

    Conclusions:

    • The optimized nOTLC-MS system successfully balances throughput and sensitivity for picogram-scale proteomics.
    • The low-loss injection method significantly enhances LC performance for limited sample analysis.
    • This platform offers a transformative solution for large-scale exploration of cellular heterogeneity with high speed and sensitivity.