title: Combination of the 2A/furin technology with an animal component free cell line development platform process.
abstract: The recently described 2A/furin technology combines both chains of the antibody in a single open reading frame. Upon translation and secretion, the peptide is processed by the cell to generate native fully functional IgG antibodies. Here, we describe the results of an evaluation study of this technology for an industrial CHO cell line development process. The 2A/furin expression cassette setup was combined with a Novartis vector system. A transfection, selection, and cloning procedure in chemically defined media was established at Novartis and applied for a monoclonal test antibody. The productivity of 2A/furin-vector-derived clones in non-optimized generic shake flask fed-batch models was in a comparable range with clones derived from the reference control vector. Higher clonal production stability was seen for the majority of clones generated with the 2A/furin technology compared to the clones generated with the reference control vector. Product quality was analyzed by SDS-PAGE and no significant difference was detected between the two systems. Thus, it was shown that the 2A/furin technology can be successfully combined with a Novartis CHO expression system and platform. Due to the single ORF setup, the 2A/furin technology may therefore offer a suitable approach to reduce vector size and complexity.
authors.affiliation: Process Sciences and Production, Novartis Biologics, Basel, Switzerland. thomas.jostock@novartis.com
authors.forename: Thomas
authors.initials: T
authors.lastname: Jostock
authors.forename: Zorica
authors.initials: Z
authors.lastname: Dragic
authors.forename: Jianmin
authors.initials: J
authors.lastname: Fang
authors.forename: Karin
authors.initials: K
authors.lastname: Jooss
authors.forename: Burkhard
authors.initials: B
authors.lastname: Wilms
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Knopf
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D045683:Furin / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism
chemical_list: D045683:Furin