Affinity chromatography of soluble fibrin in human plasma on insolubilized fibrinogen and on fragment D from fibrinogen, non cross-linked or cross-linked fibrin

Abstract

Normal human plasma containing approximately 2.5 mg of fibrinogen per ml, 131l-labelled fibrinogen (less than 0.1 mg per ml) and 125l-labelled fibrin monomer (between 0.012 and 0.05 mg per ml) was subjected to affinity chromatography on insolubilized human fibrinogen (Fg-agarose), fragment D prepared from fibrinogen (FgD-agarose), fragment D from non cross-linked fibrin (FbD-agarose) or fragment D from cross-linked fibrin (D-D-agarose). The fibrin monomer was obtained by dissolving fibrin in 2 M NaBr. The fibrinogen and fragments D were highly purified materials which were coupled to CNBr-activated Sepharose in comparable molar amounts (8 mg per ml gel for the fragments D and 20 mg per ml gel for fibrinogen). The FgD- and D-D-columns had negligible affinity for fibrinogen in plasma, the Fg-column bound approximately 3 percent of the Fibrinogen and the FbD-column 1.5 percent. All columns showed very similar binding properties for soluble fibrin monomer both with respect to the amounts of fibrin adsorbed and the amounts eluted with 2 M NaBr. Thus the reported higher affinity of insolubilized fibrin monomer to fragment D prepared from fibrin compared to Fragment D from fibrinogen was not associated with a higher affinity of insolubilized fragments D from fibrin for soluble fibrin monomer.