GPDPLQ1237 -A Type II Collagen Neo-Epitope Biomarker of Osteoclast- and Inflammation-Derived Cartilage Degradation in vitro

Scientific Reports
Henrik LöfvallKim Henriksen

Abstract

C-telopeptide of type II collagen (CTX-II) has been shown to be a highly relevant biomarker of cartilage degradation in human rheumatic diseases, if measured in synovial fluid or urine. However, serum or plasma CTX-II have not been demonstrated to have any clinical utility to date. Here, we describe the GPDPLQ1237 ELISA which targets the EKGPDPLQ↓ neo-epitope, an elongated version of the CTX-II neo-epitope (EKGPDP↓), speculated to be a blood-precursor of CTX-II generated by the cysteine protease cathepsin K. Human osteoclast cartilage resorption cultures as well as oncostatin M and tumour necrosis factor α-stimulated bovine cartilage explant cultures were used to validate GPDPLQ1237 biologically by treating the cultures with the cysteine protease inhibitor E-64 and/or the matrix metalloproteinase (MMP) inhibitor GM6001 to assess the potential contributions of these two protease classes to GPDPLQ1237 release. Cartilage resorption-derived GPDPLQ1237 release was inhibited by E-64 (72.1% inhibition), GM6001 (75.5%), and E-64/GM6001 (91.5%), whereas CTX-II release was inhibited by GM6001 (87.0%) but not by E-64 (5.5%). Cartilage explant GPDPLQ1237 and CTX-II release were both fully inhibited by GM6001 but were not inhibited by E-64....Continue Reading

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Citations

May 1, 2021·Biomolecules·Katarzyna Winsz-SzczotkaKrystyna Olczyk

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Datasets Mentioned

BETA
GM6001

Methods Mentioned

BETA
urine sampling
lavage
enzyme-linked immunosorbent assay
ELISA
biopsies
chips
ELISAs
biopsy

Software Mentioned

SPSS Statistics
SoftMax Pro
Olympus cellSens
GraphPad Prism
PattInProt
Network Protein Sequence Analysis PattInProt search 66
Fiji 73
GraphPad
NPS

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