Collagen type II C-telopeptide fragments as an index of cartilage degradation.
We report the development of an assay for measurement of the urinary concentration of collagen type II C-telopeptide fragments. This assay was developed for providing a specific marker of joint metabolism. A monoclonal antibody, recognizing a linear six amino acid epitope from the middle region of the collagen type II C-telopeptide was used in a competitive enzyme-linked immunoassay (ELISA) format for measurement of urine samples. The technical performance and specificity of the assay was evaluated and a panel of samples from patients with rheumatoid arthritis (RA) (n = 27), osteoarthritis (OA) (n = 29), Paget's disease (n = 9), and healthy controls (n = 428) was measured in the assay. The ELISA was specific for the peptide EKGPDP derived from collagen type II C-telopeptide: it did not recognize peptides from the N-telopeptide of the molecule or from other collagen types. Collagen type II C-telopeptide fragments measured in the assay resisted seven freeze-thaw cycles and >20 h of storage at room temperature. RA and OA patients showed significant 2.33-fold (95% confidence interval [CI] 1.50-3.16) and 1.53-fold (CI 1.24-1.82) elevations in CartiLaps concentration, respectively. Paget's disease patients did not have elevated CartiLaps levels. RA patients with radiological evidence of cartilage damage had significantly higher (1.79-fold, CI 1.04-2.54) CartiLaps levels than RA patients without radiological evidence of cartilage destruction. The Cartilaps assay showed high technical precision and an ability to differentiate populations with an elevated joint metabolism from normal controls. This suggests that the assay may have clinical value in assisting in the diagnosis of joint diseases and in monitoring progression and therapy in RA and OA.
Christgau
S
S
Osteometer BioTech A/S, Herlev, Denmark. sc@osteobio.com
Garnero
P
P
Fledelius
C
C
Moniz
C
C
Ensig
M
M
Gineyts
E
E
Rosenquist
C
C
Qvist
P
P
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001172:Arthritis, Rheumatoid / Q000473:pathology / Q000652:urine*; D015415:Biomarkers; D002356:Cartilage / Q000473:pathology*; D002478:Cells, Cultured; D002940:Circadian Rhythm; D003094:Collagen / Q000032:analysis / Q000276:immunology / Q000652:urine; D024042:Collagen Type I; D024043:Collagen Type II / Q000032:analysis* / Q000276:immunology / Q000652:urine; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D010001:Osteitis Deformans / Q000473:pathology / Q000652:urine; D010003:Osteoarthritis / Q000473:pathology / Q000652:urine*; D010010:Osteoclasts / Q000737:chemistry / Q000166:cytology; D010446:Peptide Fragments / Q000032:analysis / Q000276:immunology / Q000652:urine; D010455:Peptides / Q000032:analysis / Q000276:immunology / Q000652:urine; D011817:Rabbits
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D024042:Collagen Type I; D024043:Collagen Type II; D010446:Peptide Fragments; D010455:Peptides; C052929:collagen type I trimeric cross-linked peptide; D003094:Collagen