Collect. Czech. Chem. Commun. 1994, 59, 2331-2342

Microplate Assay for Hyaluronan Useful in Clinical Practice

Mária Stančíková, Vladimír Kéry, Eduard Orviský and Lívia Cibírová

Research Institute of Rheumatic Diseases, 921 01 Piešťany, Slovak Republic


A microplate assay for determination of hyaluronan (HA) has been developed. In principle, HA in the sample and biotinylated HA compete for binding to hyaluronan binding protein (HABP) coated to the plate. The amount of the microplate-bound biotinylated HA is quantified with avidin peroxidase complex. This assay is useful for 10 - 500 μg/l with < 2% relative error with 50% inhibition at 80 μg/l. The assay is simple, sensitive and particularly suitable for the determination of HA in serum and other biological fluids. The assay correlated with only commercially available radioisotope kit, Pharmacia HA-50, with correlation coefficient 0.92 at p < 0.001. Serum HA levels of patients with osteoarthritis, rheumatoid arthritis and healthy donors determined by this assay correspond to those previously determined by different methods. These results suggest that this assay is suitable for clinical use.