Biological and physical properties of fibronectin pasteurized in the presence of stabilizers.

Interest in human plasma fibronectin (Fn) as a potential clinical product for replacement therapy in septic patients has prompted the search for stabilizers to protect the protein from heat denaturation during pasteurization designed to inactivate hepatitis viruses. Fn was pasteurized (60 degrees C, 10 h) in the presence of either citrate, tricarballylate, sucrose or four mixtures of lysine, glucarate, gluconate or citrate which had been found to increase the denaturation temperature of Fn by greater than or equal to 19 degrees C. All but a citrate/gluconate mixture were effective in preventing aggregation as measured by dye fluorescence, light scattering, gel filtration and electrophoresis. Binding to gelatin was retained and immunological activity was only slightly diminished compared to a sample heated without stabilizers. Opsonic activity was measured as ability to mediate the uptake of 125I-gelatin-coated polystyrene beads by attached human monocytes. Fn heated without stabilizers underwent a transient increase in activity which was traced to formation of aggregates having elevated specific activities. Pasteurized samples had slightly elevated opsonic activities with no detectable aggregates present, while the unstabilized control was inactive. These results indicate that the physical properties of Fn as well as the functional activities of the gelatin- and cell-binding domains can be protected against thermal denaturation by various compounds.