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Harmonized Peptide Libraries Enable Practical Biofluid Selection for Developing Biomarker Assays.

Katelyn B Brusach1, Ariana E Shannon2, Alex W Joyce2

  • 1Department of Veterinary Clinical Sciences, The Ohio State University College of Veterinary Medicine, Columbus, Ohio, USA.

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|October 14, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a mass spectrometry workflow to compare proteins in feline urine, plasma, and serum. Results show urine can sometimes replace blood biofluids for protein biomarker discovery, avoiding venipuncture.

Keywords:
biofluidsbiomarkerscatsfelinemass spectrometryparallel reaction monitoringplasmaproteomicsserumurine

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Area of Science:

  • Biochemistry
  • Proteomics
  • Clinical Chemistry

Background:

  • Selecting the optimal biofluid is crucial for accurate biomarker assessment in clinical assays.
  • Identifying the most appropriate biofluid for candidate biomarker discovery can be challenging.
  • Existing methods may lack standardized approaches for cross-biofluid biomarker analysis.

Purpose of the Study:

  • To develop and validate a mass spectrometry-based workflow for parallel processing of urine, plasma, and serum samples.
  • To create harmonized peptide libraries for consistent monitoring and cross-fluid normalization.
  • To establish a reference dataset (CATalog) for guiding biofluid selection based on protein abundance in healthy felines.

Main Methods:

  • Parallel processing of paired feline urine, plasma, and serum samples using mass spectrometry.
  • Development of biofluid-specific peptide libraries and harmonization for consistent peptide monitoring.
  • Creation of the CATalog database for protein relative abundance analysis across biofluids.

Main Results:

  • Demonstrated a workflow enabling quantitative comparisons and cross-fluid normalization of peptides.
  • The CATalog dataset provides relative protein abundance data for feline urine, plasma, and serum.
  • Explored the interchangeability of blood and urinary proteins, considering sample processing and quantification.

Conclusions:

  • Urine can potentially serve as a substitute for blood biofluids in certain protein biomarker applications.
  • The developed workflow and CATalog database facilitate informed biofluid selection for feline biomarker discovery.
  • Proper sample processing is key to realizing urine's potential as an alternative to venipuncture for protein analysis.