Purification of Adenosine Deaminase from Human Red Blood Cells

Abstract

Adenosine deaminase (adenosine aminohydrolase, EC 3.5.4.4) has been purified in high yield from human erythrocytes using a simple procedure based on two chromatographic steps. In the first step enzyme is bound to a diaminohexane-Sepharose column and eluted with salt. The second step is column chromatography on Separose-bound purine riboside prepared by cross-linking the riboside to the matrix with 1,4-butanediol-diglycidyl ether. The overall purification factor was approximately 750,000 and the purity was at least 96% The procedure is suitable for small sample sizes and for purification of the variant forms of the enzyme obtained from a single individual.