ABSTRACT
Human, rat and bovine fibrinogen were radioiodinated using a modified chloramine-T method under mild labelling conditions and with a satisfactory labelling efficiency. The 125I-human, rat and bovine fibrinogen preparations obtained met the requirements with respect to clottability, low content of 125Iodide as well as stability in vitro and in vivo. SDS-PAGE mobility demonstrated that the integrity of the fibrinogen molecule was not affected by the labelling procedure.
Subject(s)
Fibrinogen/analysis , Animals , Cattle , Chromatography, Thin Layer , Electrophoresis, Polyacrylamide Gel , Humans , Iodine Radioisotopes , Isotope Labeling , Rats , Rats, Inbred StrainsABSTRACT
For the first time, the fibrinogen preparation according to Blombäck was used to purify rat fibrinogen. Intermediate as well as final products were compared with those of human fibrinogen. Yields of final products were about 25% of plasma fibrinogen. Thrombin clottability amounted to 96.5 +/- 1.3% for rat fibrinogen and 95.0 +/- 1.1% for human fibrinogen, respectively. The integrity and the characteristic differences in molecular mass of rat and human fibrinogen chains were demonstrated by means of SDS-PAGE.
Subject(s)
Fibrinogen/isolation & purification , Animals , Blood Proteins/analysis , Electrophoresis, Polyacrylamide Gel , Humans , Rats , Species Specificity , Thrombin/analysisABSTRACT
The modified chloramine-T method used for radioiodination of human, rat, and bovine fibrinogen guaranteed mild labelling conditions and a satisfactory labelling efficiency. The quality of the 125I-human, rat, and bovine fibrinogen preparations obtained met the requirements with respect to clottability, low content of 125Iodide as well as in vitro and in vivo stability.
Subject(s)
Fibrinogen/standards , Quality Control , Tosyl Compounds , Animals , Cattle , Chloramines , Electrophoresis, Polyacrylamide Gel , Fibrinogen/isolation & purification , Fibrinogen/pharmacokinetics , Humans , Iodine Radioisotopes/isolation & purification , Iodine Radioisotopes/standards , Rats , Species Specificity , Thrombin TimeABSTRACT
Hirudin was 125I-labelled using a modified chloramine-T method. 125I-hirudin proved to be a suitable marker in pharmacokinetic studies, if unchanged 125I-hirudin in body fluids was determined by means of a binding assay using immobilized thrombin. In rats and dogs a study was performed on the pharmacokinetic behaviour of hirudin following intravenous and subcutaneous injection, resp., or one-hour infusion and pharmacokinetic parameters were calculated.
