Subject(s)
Hirudins/pharmacokinetics , Macaca mulatta/blood , Thrombin/pharmacokinetics , Amino Acid Sequence , Animals , Hirudins/pharmacology , Hirudins/urine , Humans , Macaca mulatta/urine , Metabolic Clearance Rate , Molecular Sequence Data , Partial Thromboplastin Time , Species Specificity , Thrombin/pharmacology , Thrombin/urine , Thrombin TimeABSTRACT
rDNA hirudin plasma concentrations in man and rhesus monkeys were determined over a period of 15 and 24 h. The plasma concentration of alpha-human thrombin-hirudin complex was measured after administration of the complex to rhesus monkeys. The complex was also determined after administration of hirudin to man and rhesus monkeys to study a possible formation of a complex with alpha-human thrombin in blood. The determination of hirudin was performed by a sandwich ELISA, using polyclonal and monoclonal antibodies and the chromogenic thrombin substrate assay. The alpha-human thrombin-hirudin complex concentration in the plasma of rhesus monkeys was measured over a period of 48 hours. The results of a sandwich ELISA were compared with those of the chromogenic thrombin substrate assay. A good agreement between the total hirudin concentrations analyzed by the hirudin ELISA and the alpha-human thrombin-hirudin complex ELISA and those of the chromogenic thrombin substrate assay, measuring total hirudin, too, was observed.
Subject(s)
Chromogenic Compounds/analysis , DNA, Ribosomal/blood , Hirudins/blood , Thrombin/analysis , Animals , Enzyme-Linked Immunosorbent Assay , Hirudins/chemistry , Humans , Macaca mulatta , Thrombin/chemistryABSTRACT
The effects of sample preparations by dialysis and gel filtration on the catalytic concentrations of alanine aminopeptidase, N-acetyl-beta-D-glucosaminidase, and beta-glucuronidase are described. Individual urines were collected during 24 hours on 3 consecutive days from 10 male rats. Gel filtration (Sephadex G25) was more effective than dialysis against water in the removal of inhibitors of N-acetyl-beta-D-glucosaminidase and beta-glucuronidase. For alanine aminopeptidase, slightly higher results were obtained by dialysis. Inhibitor contents varied from day to day. Activity decreases of beta-glucuronidase and N-acetyl-beta-D-glucosaminidase were found in some of the urine samples and interpreted as removal of activators. Gel filtration is recommended for the preparation of rat urine for the measurement of these three enzymes. The slightly inferior effect of gel filtration on alanine aminopeptidase should be disregarded for the sake of practicality.
