Purification of uricase by biospecific adsorption-desorption.
A simple procedure for the purification of uricase from bovine kidney is described. The procedure involves the following steps: 1) processing of kidney mince by borate/butanol, 2) ammonium sulphate precipitation, and 3) biospecific adsorption-desorption. The adsorbents were prepared by chemical attachment of urate or xanthine to agarose gel beads. The desorption was performed by a xanthine solution. The adsorption-desorption procedure resulted in an 11 000-12 000-fold purification. The specific activity of the purified uricase was 19.8 U/mg using either "urate" adsorbent. The recovery was about 70%. The adsorbents were also used for the purification of commercial uricase preparations from hog liver. In this case the purified uricase also possessed a specific activity of 19.8 U/mg. The products were homogenous as judged by gradipore electrophoresis and gel filtration.
D000327:Adsorption; D000818:Animals; D002417:Cattle; D007668:Kidney / Q000201:enzymology; D012685:Sepharose; D014503:Urate Oxidase / Q000302:isolation & purification*
D012685:Sepharose; D014503:Urate Oxidase