The assessment of cartilage degradation in vivo: development of an immunoassay for the measurement in body fluids of type II collagen cleaved by collagenases.
A monoclonal antibody has been developed which recognizes a neoepitope in type II collagen which is generated by the intrahelical cleavage of collagenases. Antibody reactivity is directed at the carboxyl-terminus of the TCA or 3/4 piece of the degraded alpha1(II) chain. Reactivity is dependent upon hydroxylation of proline. Evidence is provided suggesting that epitope binding involves the recognition of a conformational neoepitope. Using an ELISA, we show that this neoepitope can be detected in the urines and sera of nonarthritic persons and patients with rheumatoid arthritis (RA). An increased content is observed in the sera and urines of patients. The assay may be of value in studying cartilage type II degradation both in vitro and in vivo such as in those with arthritis.
Poole
A Robin
AR
Joint Diseases Laboratory, Shriners Hospitals for Children, Montreal, Quebec, Canada. rpoole@shriners.mcgill.ca
Ionescu
Mirela
M
Fitzcharles
M A
MA
Billinghurst
R Clark
RC
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D001172:Arthritis, Rheumatoid / Q000097:blood* / Q000652:urine*; D024043:Collagen Type II / Q000097:blood* / Q000276:immunology / Q000652:urine*; D017364:Collagenases / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D018604:Epitope Mapping; D000939:Epitopes / Q000097:blood / Q000276:immunology / Q000652:urine; D006801:Humans; D011487:Protein Conformation; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000906:Antibodies; D024043:Collagen Type II; D000939:Epitopes; D017364:Collagenases