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Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule.
Ahmadian, Haleh; Hansen, Ernst B; Faber, Johan H; Sejergaard, Lars; Karlsson, Johan; Bolt, Gert; Hansen, Jens J; Thim, Lars.
Afiliación
  • Ahmadian H; aNovo Nordisk A/S, Biopharmaceutical Research Unit, Mål⊘v bNovo Nordisk A/S, CMC Supply, Gentofte, Denmark.
Blood Coagul Fibrinolysis ; 27(5): 568-75, 2016 Jul.
Article en En | MEDLINE | ID: mdl-26761578
Turoctocog alfa (NovoEight) is a third-generation recombinant factor VIII (rFVIII) with a truncated B-domain that is manufactured in Chinese hamster ovary cells. No human or animal-derived materials are used in the process. The aim of this study is to describe the molecular design and purification process for turoctocog alfa. A five-step purification process is applied to turoctocog alfa: protein capture on mixed-mode resin; immunoaffinity chromatography using a unique, recombinantly produced anti-FVIII mAb; anion exchange chromatography; nanofiltration and size exclusion chromatography. This process enabled reduction of impurities such as host cell proteins (HCPs) and high molecular weight proteins (HMWPs) to a very low level. The immunoaffinity step is very important for the removal of FVIII-related degradation products. Manufacturing scale data shown in this article confirmed the robustness of the purification process and a reliable and consistent reduction of the impurities. The contribution of each step to the final product purity is described and shown for three manufacturing batches. Turoctocog alfa, a third-generation B-domain truncated rFVIII product is manufactured in Chinese hamster ovary cells without the use of animal or human-derived proteins. The five-step purification process results in a homogenous, highly purified rFVIII product.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Factor VIII / Expresión Génica / Secuencia de Aminoácidos / Eliminación de Secuencia Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Blood Coagul Fibrinolysis Asunto de la revista: ANGIOLOGIA / HEMATOLOGIA Año: 2016 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Factor VIII / Expresión Génica / Secuencia de Aminoácidos / Eliminación de Secuencia Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Blood Coagul Fibrinolysis Asunto de la revista: ANGIOLOGIA / HEMATOLOGIA Año: 2016 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Reino Unido