Purification of human adenosine deaminase for the preparation of a reference material

The goal was to optimise a purification procedure of adenosine deaminase from human erythrocytes for the preparation of a European Reference Material. Adenosine deaminase was purified from human erythrocytes with a specific activity of 4.46 microkat/mg of protein and a catalytic concentration of 133 microkat/l. The isolation and purification procedure involved ion-exchange chromatography (STREAMLINE DEAE), and two purine riboside affinity chromatographies. The purified enzyme exhibits a single band in SDS-PAGE with a molecular weight of 41600 g/mol, and three bands in PAGE, isoelectric focusing and two-dimensional electrophoresis with pI 4.7, 4.85 and 5.0.