title: Overexpression of taurine transporter in Chinese hamster ovary cells can enhance cell viability and product yield, while promoting glutamine consumption.
abstract: Transporters mediate the uptake of nutrients such as amino acids and the excretion of metabolites. The fact that transporters play crucial roles in regulating cell metabolism suggests that they might be useful targets for cell engineering to enhance the yield and/or quality of monoclonal antibody (MAb) produced by CHO cells. The taurine transporter (TAUT) is stably expressed in CHO-DXB11 cells and is upregulated late in the culture period. We found that forcing the overexpression of TAUT delayed apoptotic cell death, extending the culture period. Thus, under fed-batch small-culture conditions, CHO cells that expressed pHyg-TAUT plasmid (TAUT/CHO cells), but not those that contained the null plasmid pHyg (HYG/CHO cells), produced more MAb (P < 0.01) and less lactate (P < 0.05). In a 1-L bioreactor, a representative high-yield TAUT/CHO cell line (T10) showed >80% viability for more than 1 month and a 47% increase in medium MAb concentration. In T10 cells, the upregulation of TNF-α mRNA (an apoptosis marker) and the accumulation of ammonia late in the culture period were suppressed. Moreover, if an excess of taurine was added, T10 cells efficiently consumed glutamine but not other amino acids, so T10 cells may have gained a glutamine transporter-like function. Because a considerable amount of metabolic energy is derived from glutamine, this active glutamine consumption in T10 cells might be a reason for the improved cell viability and MAb concentration. These results demonstrate that forcing the overexpression of TAUT in CHO cells can enhance cell culture performance and increase MAb titer.
authors.affiliation: Chugai Pharmaceutical Co., Ltd., Kita-ku, Tokyo, Japan. tabuchihsh@chugai-pharm.co.jp
authors.forename: Hisahiro
authors.initials: H
authors.lastname: Tabuchi
authors.forename: Tomoya
authors.initials: T
authors.lastname: Sugiyama
authors.forename: Saeko
authors.initials: S
authors.lastname: Tanaka
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tainaka
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D005973:Glutamine / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D026901:Membrane Transport Proteins / Q000096:biosynthesis*; D000099247:Taurine Transporters
chemical_list: D005973:Glutamine; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D000099247:Taurine Transporters