Removal of protease from Streptomyces hyaluronidase by affinity chromatography

A simple method is presented for removing protease(s) from commercial sources of Streptomyces hyaluronidase derived from Streptomyces hyalurolyticus using a column of insolubilized chicken ovomucoid. The protease binds to ovomucoid; 91% of the hyaluronidase and no more than 0.7% of the protease passes through the column. A calcium chloride solution is used to remove the bound protease. Purified hyaluronidase cleaves the hyaluronic acid of proteoglycan aggregates without affecting the size of proteoglycan monomers. The ability of digested monomers to bind hyaluronic acid, however, is somewhat diminished.