Adsorption and function of recombinant Factor VIII at the air-water interface in the presence of Tween 80.

The air-water surface tension kinetic and steady state behavior exhibited by a recombinant Factor VIII were recorded in the presence of the surfactant Tween 80. The rate of surface tension decrease was determined to be greater for rFVIII-Tween mixtures than for Tween acting alone, at all Tween concentrations studied (8-108 ppm). In addition, steady state interfacial behavior was determined to be governed entirely by surfactant adsorption in mixtures with Tween concentrations greater than or equal to about 18 ppm. The formation of aggregates, tertiary structure change, and biological activity of rFVIII in agitated as well as unagitated samples were monitored using turbidity measurements, intrinsic fluorescence spectroscopy, and the activated partial thromboplastin time, respectively. The results are consistent with two mechanisms characterizing the surfactant-mediated stabilization of rFVIII: the rapid, preferential adsorption of Tween molecules relative to rFVIII molecules at the air-water interface, and Tween association with rFVIII in solution to provide a steric barrier to aggregation. While both mechanisms are relevant in the presence of an interface, evidence recorded here suggests that the preferential adsorption of Tween at the interface is the major stabilization mechanism while the effectiveness of Tween association with rFVIII is compromised in agitated samples.