Accumulation of hyaluronate in human lung carcinoma as measured by a new hyaluronate ELISA.
We have developed a new ELISA to quantify hyaluronate. This ELISA takes advantage of an anti-keratan sulfate antibody to differentiate between the coated aggregating rat chondrosarcoma proteoglycan which captures the hyaluronate and the keratan sulfate-bearing aggregating proteoglycan added subsequently. Absorbance in this assay was shown to be linear to the logarithmic concentration of hyaluronate in the range of 15 to 1000 ng/ml. Pre-treatment of hyaluronate with papain or protease did not interfere with its quantification; in contrast, pre-treatment with 0.1N NaOH significantly interferes with the subsequent measurement of the hyaluronate molecules. The size of the hyaluronate molecule was found to be an important factor in quantification: only large size hyaluronate molecules could be quantified accurately. The ELISA was used to show that human lung carcinomas contain 2 to 500 times as much hyaluronate as normal lung tissue from the same patient.
Li
X Q
XQ
Department of Biochemistry, Rush-Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612.
Thonar
E J
EJ
Knudson
W
W
D000911:Antibodies, Monoclonal; D002277:Carcinoma / Q000032:analysis*; D011508:Chondroitin Sulfate Proteoglycans / Q000276:immunology; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation*; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007632:Keratan Sulfate / Q000276:immunology; D000071234:Lumican; D008168:Lung / Q000032:analysis; D008175:Lung Neoplasms / Q000032:analysis*; D008970:Molecular Weight
D000911:Antibodies, Monoclonal; D011508:Chondroitin Sulfate Proteoglycans; C000604306:LUM protein, human; C000604308:Lum protein, rat; D000071234:Lumican; D006820:Hyaluronic Acid; D007632:Keratan Sulfate