Molecular Origins of Recombinant Factor VIII (rFVIII) Public Deposited

http://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/z890rw203

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  • The capability of Pluronic® F68 to stabilize human recombinant Factor VIII in solution was assessed using zeta potential measurements. Hemophilia A is a genetic disorder in which the body is missing Factor VIII (FVIII) from the blood coagulation cascade. The pharmaceutical industry developed recombinant Factor VIII (rFVIII), the largest cloned molecule, as a viable treatment option for the missing protein. The protein suffers instability in solution which leads to aggregation, precipitation, and adsorption. Current formulations of rFVIII include the surfactant Tween 80 as a stabilization aid. Pluronic® F68, another surfactant, has been shown to prevent sheer in mammalian cell cultures mainly by enhancing cell resistance to sheer stress. Pluronic® F68 may be able to increase the stability of rFVIII for prevention of aggregation and precipitation. The stabilization of rFVIII was monitored using a zeta potentiometer with silica as a model surface. Zeta potential was unable to register a significant change for the initial concentrations of rFVIII used. Investigations were performed by increasing the amount of rFVIII and testing the effect of Tween 80 on the silica microspheres. The zeta potentiometer was not sensitive enough to register any conclusive results as to the effectiveness of Pluronic® F68 due to interactions with Tween 80. Further research is required to determine the effectiveness of Pluronic® F68 for rFVIII stabilization.
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