Ethanolic fractionation of bovine testicular hyaluronidase.

Studies on the fractionation of blood serum by Cohn et al. (1) have demonstrated that under precisely controlled conditions reproducible fractionation of a complex mixture of proteins can be accomplished by the use of ethanol. This paper reports the application of this method to the separation of hyaluronidase from testicular extract. A variety of experimental conditions involving variation of pH, ionic strength, and protein concentration was employed in order to ascertain optimum conditions for purification. Enzyme purity was followed in terms of activity per mg. of nitrogen.