title: Simultaneous targeting of Requiem & Alg-2 in Chinese hamster ovary cells for improved recombinant protein production.
abstract: Apoptosis is known to be the main cause of cell death in the bioreactor environment, leading to the loss of recombinant protein productivity. In a previous study, transcriptional profiling was used to identify and target four early apoptosis-signaling genes: FADD, FAIM, Alg-2, and Requiem. The resulting cell lines had increased viable cell numbers and extended culture viability, which translated to increased protein productivity. Combinatorial targeting of two genes simultaneously has previously been shown to be more effective than targeting one gene alone. In this study, we sought to determine if targeting Requiem and Alg-2 was more effective than targeting Requiem alone. We found that targeting Requiem and Alg-2 did not result in extended culture viability, but resulted in an increase in maximum viable cell numbers and cumulative IVCD under fed-batch conditions. This in turn led to an approximately 1.5-fold increase in recombinant protein productivity.
authors.affiliation: Department of Chemical Engineering and Chemical Technology, Imperial College London, London, SW7 2AZ, UK.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Lim
authors.forename: Athanasios
authors.initials: A
authors.lastname: Mantalaris
authors.forename: Miranda G S
authors.initials: MG
authors.lastname: Yap
authors.forename: Danny C F
authors.initials: DC
authors.lastname: Wong
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics*; D001483:Base Sequence; D016466:CHO Cells; D002135:Calcium-Binding Proteins / Q000235:genetics*; D020169:Caspases / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics*; D006868:Hydrolysis; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors
chemical_list: D051017:Apoptosis Regulatory Proteins; D002135:Calcium-Binding Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; C113581:DPF2 protein, human; C097753:PDCD6 protein, human; D011994:Recombinant Proteins; D014157:Transcription Factors; D007371:Interferon-gamma; D020169:Caspases