Molecular origins of surfactant stabilization of a human recombinant factor VIII Public Deposited

http://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/n296x055s

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  • The project objective was to characterize stabilization of human recombinant Factor VIII (FVIII) with two surfactants, Pluronic® F-68 (F68) and Polysorbate 80 (PS80). Factor VIII is an essential blood clotting protein manufactured for treatment of Hemophilia A. Factor VIII has multiple hydrophobic regions requiring the use of amphiphilic molecules, such as surfactants, to stabilize the protein in aqueous solutions. Over half of FVIII produced is lost due to adsorption and aggregation during current downstream processing using PS80. Du Nouy ring interfacial tensiometry was used to measure surface tension depression of surfactant samples ranging from 10-150 ppm with and without 60-70 IU/mL FVIII injected into KG-2 buffer at temperatures ranging from 4 to 60 °C. Addition of FVIII caused the rate of surface tension depression to increase by 15% in PS80 and decrease by 26% in F68. Surface tension depression data indicates that PS80 preferentially binds to the air-water interface while F68 prefers to associate with FVIII, helping to stabilize the protein in solution and prevent adsorptive and aggregative losses. The results suggest that FVIII yield could be substantially improved by the replacement of PS80 with F68 in commercial production.
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  • description.provenance : Submitted by Jakson Clark (clarkjak@onid.orst.edu) on 2013-06-03T18:46:29Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: bb87e2fb4674c76d0d2e9ed07fbb9c86 (MD5) Clark_et_al_Expo.pdf: 1426393 bytes, checksum: 21336734b3354902ed1ebd57e7bd2d22 (MD5)
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  • description.provenance : Approved for entry into archive by Deanne Bruner(deanne.bruner@oregonstate.edu) on 2013-06-04T18:22:28Z (GMT) No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: bb87e2fb4674c76d0d2e9ed07fbb9c86 (MD5) Clark_et_al_Expo.pdf: 1426393 bytes, checksum: 21336734b3354902ed1ebd57e7bd2d22 (MD5)

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