Chemistry of carbon compounds – Miscellaneous organic carbon compounds – C-metal
Patent
1980-06-02
1981-07-14
Schain, Howard E.
Chemistry of carbon compounds
Miscellaneous organic carbon compounds
C-metal
424101, A23J 106, A61K 3516, C07G 700
Patent
active
042785944
ABSTRACT:
A process for the separation and isolation of AHF, fibronectin and von Willebrand's ristocetin cofactor from blood plasma by adding a sulfated mucopolysaccharide to blood plasma to a concentration of from about 0.05 to about 0.4 mg/ml of plasma, cooling the plasma to 0.degree. C.-15.degree. C., centrifuging the plasma to isolate a precipitate which forms and then removing the AHF-rich supernatant liquid from the precipitate.
The precipitate is washed in a slightly basic solution of dilute salts and then dissolved in a buffered solution of chaotrophic salts of the Hofmeister series having an ionic strength from 0.01-1. Fibronectin is separated by chromotographic means from the dissolved precipitate which contains fibronectin and fibrinogen.
The dissolved precipitate may also be applied to a glass or plastic column containing a 4% agarose gel (E. G. Sepharose 4B or Biogel A-15m) in a 0.25 molar solution sodium phosphate titrated to pH 7.0 with any base, preferably TRIS. The von Willebrand's ristocetin cofactor elutes in the void volume of the column, whereas the fibronectin-rich fraction elutes later. Both fractions may be collected and concentrated.
in a preferred embodiment, 0.20 mg of sodium heparin per milliliter of human plasma is employed with the heparinized plasma solution cooled for 3 hours at a temperature of 2.degree. C.-4.degree. C.
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