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Clinical Chemistry 19: 908-918, 1973;
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Clinical Chemistry, Vol 19, 908-918, Copyright © 1973 by the American Association for Clinical Chemistry

Evaluation of Kinetic Enzyme Parameters by Use of a Small Computer Interfaced "Fast Analyzer"—An Addition to Automated Clinical Enzymology

T. O. Tiffany 1, D. D. Chilcote 1, and C. A. Burtis 1

1 Molecular Anatomy (MAN) Program,1 Oak Ridge National Laboratory,2 Oak Ridge, Tenn. 37830.

The evaluation of steady-state enzyme kinetic parameters such as Km, Vmax, and Ki is useful in clinical enzymology, particularly where they can be used for the detection of enzyme variants and to assess the type of isoenzymes and the extent of their abnormal activity. The multiple-cuvet parallel-analysis feature of the Fast Analyzer permits one to evaluate one or more enzymic parameters in a single rotor. Both on-line measurement of initial reaction velocities and rapid off-line reduction of statistical data can be obtained by using the small-computer interfaced Fast Analyzer. Estimates of error in determining these parameters are also obtained, for use in comparing results in future analyses. We discuss the programming approach for adapting both simple and more complex statistical nonlinear regression data-fitting routines to the Fast Analyzer’s computer. We illustrate how the analyzer may be used to determine simple kinetic parameters for placental alkaline phosphatase and glucose-6-phosphate dehydrogenase and for the more complex "stripping out" of Km’s and Vmax’s for a heterogenous mixture of placental and intestinal alkaline phosphatase.


Key Words: alkaline phosphatase • glucose-6-phosphate dehydrogenase • isoenzymes • Km, Vmax, Ki evaluations • enzyme kinetics • FOCAL • computer programming • "isoenzyme stripping"

Submitted on May 3, 1973
Accepted on May 29, 1973






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Copyright © 1973 by the American Association for Clinical Chemistry.