Evaluation of the virus clearance capacity and robustness of the manufacturing process for the recombinant factor VIII protein, turoctocog alfa.
Turoctocog alfa is a B-domain-truncated recombinant factor VIII protein produced in a Chinese hamster ovary (CHO) cell line. The aim of this study was to evaluate the virus clearance capacity and robustness of the turoctocog alfa purification process. Virus clearance evaluation studies were conducted utilising a scaled-down version of the manufacturing process. Total virus clearance was evaluated using the ecotropic murine leukaemia virus (eMuLV) as a model for non-infectious retrovirus-like particles (RVLPs) and certain enveloped viruses. Other viruses utilised included: infectious bovine rhinotracheitis (IBRV), minute virus of mice (MVM), bovine enterovirus (BEV) and Reo-3 virus (Reo-3). Robust clearance of all model viruses was demonstrated with either new or reused resins. Overall, virus reduction factors were: >18.0 log10(eMuLV); 11.0 log10(MVM); >11.8 log10(Reo-3; >5.0 log10using nanofiltration); >15.3 log10(BEV) and >12.7 log10(IBRV). Taken together, these values demonstrate that the purification process for turoctocog alfa effectively removes a range of enveloped and non-enveloped viruses of different physicochemical properties and sizes.
Dept. 609, Biopharm Purification Development & Virology, Novo Nordisk A/S, Hagedornsvej 1, DK-2820 Gentofte, Denmark. Electronic address: tryw@novonordisk.com.
Tracey W
TW
Ellgaard
Dept. 609, Biopharm Purification Development & Virology, Novo Nordisk A/S, Hagedornsvej 1, DK-2820 Gentofte, Denmark. Electronic address: lnbi@novonordisk.com.
Lene
L
Bindslev
Dept. 609, Biopharm Purification Development & Virology, Novo Nordisk A/S, Hagedornsvej 1, DK-2820 Gentofte, Denmark. Electronic address: skam@novonordisk.com.
Søren
S
Kamstrup
D000818:Animals; D016466:CHO Cells; D002417:Cattle; D006224:Cricetinae; D003412:Cricetulus; D029650:Enterovirus, Bovine; D005169:Factor VIII / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D007242:Herpesvirus 1, Bovine; D009052:Leukemia Virus, Murine; D051379:Mice; D008915:Minute Virus of Mice; D011994:Recombinant Proteins; D038301:Virus Inactivation
D011994:Recombinant Proteins; C078147:F8 protein, human; D005169:Factor VIII
Manufacture; Turoctocog alfa; Virus clearance; Virus clearance evaluation; Virus safety; rFVIII