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Assay of plasma leupeptin using the reversible binding of leupeptin to bovine pancreatic trypsin |
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| Authors: | K Yamashita K Watanabe H Takayama S Mizuguchi M Ishibashi H Miyazaki W Tanaka H Umezawa |
| Affiliation: | 1. Office of New Drugs, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA;2. Office of Clinical Pharmacology, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA |
| Abstract: | A competitive binding radioassay for leupeptin has been developed utilizing the reversible binding of leupeptin to bovine pancreatic trypsin. An ethanol precipitation step was introduced to separate trypsin-bound leupeptin from its free form. Advantages of this method are simplicity of the procedure and avoidance of the preparation of antiserum. The possible metabolites of leupeptin exhibit no significant inhibitory effect on leupeptin-trypsin binding in this system. This method was applied to the determination of plasma leupeptin levels in dogs after oral administration of the peptide. |
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