A great new paper from our founder's lab, Dr. Michael Yu at the University of Utah! They developed a dimerized version of the CHP that is linked on the C-terminus that allows for faster-refolding kinetics which can be useful for creating ELISA assays to detect low levels of damaged collagen in biological fluids. They show that it has the ability to capture fragments of collagen that would otherwise go unnoticed and were able to facilitate early peptidomic-based analysis for a mouse osteoporosis model. This may prove helpful in peptidomic-based disease detection and biomarker discovery.
Abstract: Collagen remodeling in normal and pathologic conditions releases numerous collagen fragments into biological fluids. Although a few collagen fragments have been tested as biomarkers for disease indication, most occur at trace levels, making them nearly impossible to detect even with modern analytical tools. Here we report a new way to enrich collagen fragments that allows complete peptidomic analysis of collagen fragments in urine. Enrichment is made possible by dimeric collagen hybridizing peptides (CHPs) that bind collagen fragments originating from the triple-helical regions of all collagen types with minimal sequence bias. LC-MS/MS analysis of enriched mouse urine revealed an average of 383 collagenous peptide fragments per sample (compared to 34 for unenriched sample) which could be mapped to all types of mouse collagens in the SwissProt database including FACITs and MACITs. Hierarchical clustering of a selected panel of the detected fragments separated osteopenic mice from healthy mice. The results demonstrate dimeric CHP’s ability to enrich collagen fragments from biological fluid and its potential to aid peptidomics-based disease detection and biomarker discovery. [Raw mass spectrometry files have been deposited to the MassIVE repository (massive.ucsd.edu, MSV000085372, doi 10.25345/C5N12H)].
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