Graduate Thesis Or Dissertation
 

The adsorption of human recombinant factor VIII in the presence of the nonionic triblock surfactant Pluronic® F-68 at the air-water interface

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  • The adsorption behavior of a human recombinant Factor VIII (rFVIII) in the presence of the nonionic, poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock surfactant Pluronic® F-68 was evaluated by dynamic air-water tensiometry. Surface tension kinetics were recorded for F-68 in the absence of protein at concentrations ranging from 0 to 10,000 ppm. The rate and extent of surface tension depression in these samples was compared to dynamic tensiometry measurements recorded for F-68 solutions in the presence of rFVIII. Surfactants are used in the biopharmaceutical industry to stabilize therapeutic proteins from aggregation and adsorption loss during manufacturing, and this comparison suggested a substantially different mechanism for rFVIII stabilization by F-68 than that consistent with the kinds of surfactants commonly used in downstream processing. In particular, comparative kinetic data were consistent with the notion that with sufficient F-68 in solution, rFVIII did not adsorb at the air-water interface even though the interface was only partially occupied by F-68. We hypothesized that F-68 stabilized rFVIII in these tests by strong association with the protein itself, rather than by its adsorption and subsequent establishment of a steric barrier at the air-water interface. To test this hypothesis, turbidimetric measurements of rFVIII aggregation behavior in solution and zeta potential measurements of rFVIII adsorption on (1 μm) colloidal silica particles, each in the presence of selected concentrations of F-68, were evaluated. Results recorded with these approaches were inconclusive, but not inconsistent with the original hypothesis.
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Déclaration de droits
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