Bovine Platelet Proteins II. Purification of Platelet Fibrinogen

Summary

1. Platelet fibrinogen has been purified from washed bovine platelets. The procedure was based on the methods for purification of plasma fibrinogen by fractionated precipitations and extractions with ethanol and glycine below 0°, and precipitation of proteins by dimethylformamide at 0°.

2. The platelet extract obtained by freezing and thawing of the cells, freed from insoluble material by centrifugation at 23,000 x g for 30 min, contained 0.22 ±0.003mg fibrinogen per 10⁹ platelets. Total protein of this fraction was 0.77 ±0.08 mg per 10⁹ platelets whereas that of the insoluble fraction was 0.79 ±0.09 mg per 10⁹ platelets.

3. The most purified platelet fibrinogen fraction contained 91-98% of the protein in a thrombin-clottable state. The yield was approx. 20%. It showed homogeneity in analytical ultracentrifugation, in immunoelectrophoresis using an antiserum produced by immunization of rabbits against platelet extract, and in starch gel electrophoresis using a discontinuous system of Tris HCl and borate buffers offering a high resolution power towards the platelet proteins. Polyacrylamide disc electrophoresis revealed two to three faint lines behind the main fibrinogen line. At least one such line was also observed with purified plasma fibrinogen.

¹⁾ On WHO fellowship. Permanent adress: Institute of Haematology, Chocimska 5, Warsaw, Poland.

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