A MICROPLATE ASSAY FOR HYALURONIDASE AND HYALURONIDASE INHIBITORS

A sensitive, high-throughput assay has been developed which measures h yaluronidase activity in a microplate format. In this assay, hyaluroni c acid is suspended in agarose in a microtiter well, the plate is incu bated with the hyaluronidase solution, and the undigested hyaluronic a cid is precipitated with cetylpyridinium chloride. The precipitate blo cks light transmittance and therefore an increase in the visible light transmitted correlates with the amount of digested hyaluronic acid. U sing this assay, as little as 0.05 units of hyaluronidase activity can be detected. The assay is highly reproducible and can be run with com mercially available reagents. Hyaluronidase activity is easy to quanti tate using a microplate reader and this format allows large numbers of samples to be assayed. (C) 1994 Academic Press, Inc.