A new sandwich-ELISA method for the determination of keratan sulphate peptides in biological fluids employing a monoclonal antibody and labelled avidin biotin technique.
A new sandwich-ELISA for the determination of keratan sulphate peptides in biological fluids is described. The technique involves binding a commercially available monoclonal antibody against keratan sulphate to microtitre plates, adding the unknown keratan sulphate antigen then interacting the keratan sulphate-antibody complex with a biotin-monoclonal antibody conjugate which was also specific for keratan sulphate peptides. Alkaline phosphatase conjugated streptravidin was then added and the amount which bound to the biotin was determined by measuring the release of chromogen from an added chromogenic substrate. Using this assay, keratan sulphate peptides in biological fluids within the range 10-1000 ng/ml could be quantitated. This method was found to be more sensitive than presently used techniques. The intra- and inter-assay coefficients of variation were 11% and 13%, respectively.
Kongtawelert
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Raymond Purves Research Laboratories, (University of Sydney, Royal North Shore Hospital of Sydney, St. Leonards, N.S.W., Australia.
Ghosh
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D000328:Adult; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007632:Keratan Sulfate / Q000032:analysis* / Q000097:blood; D008875:Middle Aged; D010003:Osteoarthritis / Q000097:blood / Q000378:metabolism; D010455:Peptides / Q000032:analysis* / Q000097:blood; D011509:Proteoglycans / Q000097:blood; D013582:Synovial Fluid / Q000737:chemistry
D010455:Peptides; D011509:Proteoglycans; D007632:Keratan Sulfate