Clinical Chemistry
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Clinical Chemistry 18: 374-383, 1972;
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Clinical Chemistry, Vol 18, 374-383, Copyright © 1972 by the American Association for Clinical Chemistry

An L-Aspartate: 2-Oxoglutarate Aminotransferase Reference Material from Human Erythrocytes: Preparation and Characterization

Robert Rej 1, Raymond E. Vanderlinde 1, and Charles F. Fasce Jr. 1

1 Division of Laboratories and Research, New York State Department of Health, Albany, N.Y. 12201.

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.


Key Words: cation- and ion-exchange chromatography • reference enzyme preparation • effect of protein • Arrhenius plots • intercomparison of assays by other methods • interlaboratory comparisons

Submitted on October 6, 1971
Accepted on February 14, 1972






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Copyright © 1972 by the American Association for Clinical Chemistry.