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.
Bota
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Departament de Bioquímica i de Biologia Molecular, Unitat de Bioquímica de Medicina, Universitat Autònoma de Barcelona, Spain.
Gella
F J
FJ
Canalias
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F
D000243:Adenosine Deaminase / Q000302:isolation & purification*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007525:Isoelectric Focusing; D012015:Reference Standards
D000243:Adenosine Deaminase