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A multiplexed LC-MS/MS method to reveal changes in inflammatory and coagulation cascades induced by host infection

  • 0Department of Chemical Sciences, University of Naples Federico II, 80126, Naples, Italy.
Analytical and Bioanalytical Chemistry +

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June 25, 2025

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June 25, 2025

View abstract on PubMed

Summary

This summary is machine-generated.

This study developed a rapid mass spectrometry method to analyze proteins involved in inflammation and blood clotting, identifying key changes in COVID-19 patients for better infection monitoring.

Area Of Science

  • Proteomics
  • Clinical Chemistry
  • Immunology

Background

  • Severe respiratory infections like COVID-19 involve inflammation (cytokine storm) and coagulation abnormalities, increasing clot risk.
  • Current antibody-based tests (ELISA) for infection monitoring have limitations including complexity and cross-reactivity.
  • There is a need for advanced analytical methods to investigate host responses to infection.

Purpose Of The Study

  • To develop a mass spectrometry (MS)-based method for analyzing host response proteins in infection.
  • To establish a rapid protein digestion protocol combined with targeted tandem MS for clinical applications.
  • To quantify proteins related to inflammation and coagulation in COVID-19 patients.

Main Methods

  • Utilized S-Trap column digestion for rapid sample preparation, reducing processing time and eliminating desalting.
  • Employed a targeted tandem mass spectrometry (MS) approach with multiple reaction monitoring (MRM) ion mode.
  • Quantified approximately 60 proteins involved in inflammatory and coagulation pathways using multiplexed LC-MRM/MS on triple quadrupole mass spectrometers.

Main Results

  • The rapid digestion protocol significantly reduced sample processing time.
  • 90% of analyzed proteins showed a good instrumental response.
  • 63% of proteins displayed significant dysregulation in COVID-19 patients compared to controls, highlighting key infection-related protein changes.

Conclusions

  • The developed multiplexed LC-MRM/MS method provides high specificity and performance for clinical analysis.
  • This approach enables rapid and accurate quantification of host response proteins.
  • The method supports clinical investigations into infection pathogenesis and monitoring.

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