MSACL 2026 Abstract

From Collagen Fragments to Molecular Signatures: A Targeted LC-MS/MS Profiling Approach

Justine Demeuse (1), Lynn Kalut (1), Alix Mackowiak (1), Marine Piette (1), Elodie Grifnée (2), Etienne Cavalier (1,2)
(1) University of Liège, Liège, BE (2) University Hospital of Liège, BE

Justine Demeuse, PhD in Biomedical and pharmaceutical sciences (Presenter) University Hospital of Liège

Presenter Bio: I am a researcher in clinical mass spectrometry, focusing on the development of next-generation biomarker strategies based on targeted LC-MS/MS. I studied Biomedical Sciences at the University of Liège, where I completed both my Bachelor’s and Master’s degrees, followed by a PhD in the Faculty of Medicine within the Laboratory of Clinical Chemistry. My doctoral work focused on the characterization of type I collagen crosslinked telopeptides (CTX) using high-resolution mass spectrometry.

I am currently working at the Laboratory of Clinical Chemistry of the CIRM (University of Liège), where my research explores collagen-derived peptides as molecular signatures of tissue remodeling, with applications in musculoskeletal diseases and beyond. I am particularly interested in moving from traditional immunoassays toward more specific, multiplexed, and structurally defined approaches to support personalized medicine.

Relevant Financial Disclosures (within past 24 months, reported on Jul 06, 2026) No relevant financial relationship(s) to disclose.

INTRODUCTION:
Personalized medicine relies on the ability to stratify patients based on underlying biological mechanisms rather than treating diseases as homogeneous entities. In this context, biomarkers reflecting tissue remodeling processes are of particular interest. Collagen-derived peptides have been widely used as biomarkers across multiple diseases, including osteoporosis, osteoarthritis, cardiovascular diseases and cancer. However, these markers are almost exclusively measured using immunoassays, which suffer from well-known limitations, including limited specificity, cross-reactivity, lack of molecular definition of the measured species and poor standardization across quantitation methods.

OBJECTIVES:
To address these limitations, we developed a mass spectrometry-based “collagen profiling” approach aimed at providing a comprehensive and molecularly defined assessment of collagen turnover. As a proof of concept, we focused on collagens type I, II and III, major structural components of bone, cartilage and synovial tissues, respectively and therefore highly relevant to musculoskeletal diseases such as osteoarthritis.

METHODS:
An in silico multi-enzymatic digestion (trypsin, Glu-C and chymotrypsin) of collagen I-III sequences was performed using Skyline to generate a comprehensive list of candidate peptides and associated MRM transitions. To experimentally validate peptide detectability, purified bone, cartilage and synovial tissues were subjected to bacterial collagenase digestion followed by multi-enzymatic digestion. Peptides were fractionated by size exclusion chromatography (Superdex 30, Cytiva, ÄKTA system) and analyzed using a Sciex QTRAP 6500 coupled to a Shimadzu LC system. Chromatographic separation was achieved on an Acquity HSS T3 column using a generic gradient (H₂O/ACN, 0.4% formic acid). For each peptide, 2-3 transitions were selected based on sensitivity and specificity.

RESULTS:
This workflow enabled the detection of 132 peptides from collagen type I (47% sequence coverage), 74 from type II (49%) and 55 from type III (40%), demonstrating a high level of coverage across structurally complex proteins.

CONCLUSION:
These results highlight the feasibility of large-scale, multiplexed quantification of collagen-derived peptides using targeted LC-MS/MS.

This approach provides a significantly more specific and molecularly defined alternative to immunoassays and opens the way to multiplexed biomarker panels capturing tissue-specific remodeling processes. Ongoing work focuses on the development of sample preparation workflows, including affinity-based enrichment, to enable the application of this strategy to human plasma. Beyond musculoskeletal diseases, collagen profiling by mass spectrometry has the potential to provide clinically actionable insights across a wide range of pathologies involving extracellular matrix remodeling, supporting improved patient stratification and more personalized clinical decision-making.