Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule

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 p...

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Main Authors:	Ahmadian, Haleh, Hansen, Ernst B., Faber, Johan H., Sejergaard, Lars, Karlsson, Johan, Bolt, Gert, Hansen, Jens J., Thim, Lars
Format:	Online
Language:	English
Published:	Lippincott Williams And Wilkins 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4935534/

id	pubmed-4935534
recordtype	oai_dc
spelling	pubmed-49355342016-07-26 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 Original Articles 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. Lippincott Williams And Wilkins 2016-07 2016-01-12 /pmc/articles/PMC4935534/ /pubmed/26761578 http://dx.doi.org/10.1097/MBC.0000000000000477 Text en Copyright © 2016 Wolters Kluwer Health, Inc. All rights reserved. http://creativecommons.org/licenses/by-nc-nd/4.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License, where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially. http://creativecommons.org/licenses/by-nc-nd/4.0
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Ahmadian, Haleh Hansen, Ernst B. Faber, Johan H. Sejergaard, Lars Karlsson, Johan Bolt, Gert Hansen, Jens J. Thim, Lars
spellingShingle	Ahmadian, Haleh Hansen, Ernst B. Faber, Johan H. Sejergaard, Lars Karlsson, Johan Bolt, Gert Hansen, Jens J. Thim, Lars Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule
author_facet	Ahmadian, Haleh Hansen, Ernst B. Faber, Johan H. Sejergaard, Lars Karlsson, Johan Bolt, Gert Hansen, Jens J. Thim, Lars
author_sort	Ahmadian, Haleh
title	Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule
title_short	Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule
title_full	Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule
title_fullStr	Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule
title_full_unstemmed	Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule
title_sort	molecular design and downstream processing of turoctocog alfa (novoeight), a b-domain truncated factor viii molecule
description	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.
publisher	Lippincott Williams And Wilkins
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4935534/
_version_	1613605186898493440

Molecular design and downstream processing of turoctocog alfa (NovoEight), a B-domain truncated factor VIII molecule

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