An L-Aspartate: 2-Oxoglutarate Aminotransferase Reference Material from Human Erythrocytes: Preparation and Characterization
Open Access
- 1 April 1972
- journal article
- research article
- Published by Oxford University Press (OUP) in Clinical Chemistry
- Vol. 18 (4) , 374-383
- https://doi.org/10.1093/clinchem/18.4.374
Abstract
L-Aspartate: 2-oxoglutarate aminotransferase (EC 2.6.1.1) from human erythrocytes is purified for use as a reference material in clinical assays for the enzyme. The procedure involves heat treatment in the presence of a substrate (2-oxoglutarate), ammonium sulfate precipitation, and fractionation with n-butanol. The material is further purified by anion- and cation-exchange chromatography. Specific activity obtainable by a typical preparation is 110 µmol min-1 mg-1 at 30°C. Identical behavior is demonstrated between enzyme in human serum and purified erythrocyte enzyme when substrate concentrations, temperature, and pH are varied. A lyophilized preparation at 4°C is stable for longer than 400 days. Data regarding purity and behavior in currently used clinical assays is described.Keywords
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