ELISA for the core protein of the cartilage large aggregating proteoglycan, aggrecan: comparison with the concentrations of immunogenic keratan sulphate in synovial fluid, serum and urine.
Immunological assays for fragments of the cartilage large aggregating proteoglycan, aggrecan, have been widely used to monitor cartilage turnover. These assays have commonly employed the monoclonal keratan sulphate antibody, 5D4. Keratan sulphate, however, is present in many tissues and 5D4 affinity is critically dependent on antigen structure. We have therefore raised and characterized a monoclonal antibody (1-F21) that reacts with the core protein of aggrecan without interference from the glycosaminoglycan side chains and, using this antibody, we have optimized a sensitive, competitive ELISA. The within-assay and between-assay coefficients of variation were 4.9-8.9% and 11.1-13.0%, respectively. The mean concentrations of core protein in synovial fluid, serum and urine were 76.4 micrograms/ml, 104.0 ng/ml and 81.0 ng/ml, respectively. In synovial fluids the concentrations were closely correlated with the concentrations of keratan sulphate as determined by 5D4 (r = 0.94), whereas in serum and urine there was no obvious correlation between the determinations. These findings show that measurement of both core protein and keratan sulphate results in a more precise description of aggrecan turnover.
Department of Clinical Chemistry, K.H. University Hospital, Aarhus, Denmark.
H J
HJ
Møller
F S
FS
Larsen
T
T
Ingemann-Hansen
J H
JH
Poulsen
D053674:Aggrecans; D000818:Animals; D000911:Antibodies, Monoclonal; D002356:Cartilage / Q000737:chemistry; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis / Q000097:blood / Q000652:urine; D004797:Enzyme-Linked Immunosorbent Assay; D016326:Extracellular Matrix Proteins; D006801:Humans; D007632:Keratan Sulfate / Q000032:analysis* / Q000097:blood / Q000652:urine; D037181:Lectins, C-Type; D051379:Mice; D011506:Proteins / Q000276:immunology; D011509:Proteoglycans / Q000032:analysis* / Q000097:blood / Q000652:urine; D012680:Sensitivity and Specificity; D013582:Synovial Fluid / Q000737:chemistry*
C508320:Acan protein, mouse; D053674:Aggrecans; D000911:Antibodies, Monoclonal; D011508:Chondroitin Sulfate Proteoglycans; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011506:Proteins; D011509:Proteoglycans; D007632:Keratan Sulfate