Polyclonal antibodies against human fibroblast collagenase and the design of an enzyme-linked immunosorbent assay to measure TIMP-collagenase complex.
A polyclonal antibody has been raised against purified human fibroblast collagenase and characterised. This antibody has been used in combination with a monoclonal anti-TIMP antibody in a double antibody sandwich ELISA to measure TIMP-collagenase complex. The assay range was 5-50 ng/ml complex, quantitated in terms of the TIMP component. The assay can measure complex even in the presence of at least a 40-fold excess of free TIMP. The level of TIMP-collagenase complex has been measured in serial samples of synovial fluid from two patients with septic arthritis; high levels of complex are found in some samples, and the level of complex shows an inverse relationship with the level of free TIMP.
Clark
I M
IM
Rheumatology Research Unit, Addenbrooke's Hospital, Cambridge, UK.
Wright
J K
JK
Cawston
T E
TE
Hazleman
B L
BL
D000328:Adult; D000368:Aged; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D001170:Arthritis, Infectious / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000201:enzymology; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008297:Male; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D011817:Rabbits; D013582:Synovial Fluid / Q000737:chemistry; D019714:Tissue Inhibitor of Metalloproteinases
D000906:Antibodies; D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D019714:Tissue Inhibitor of Metalloproteinases; D003012:Microbial Collagenase