Development of transfection and high-producer screening protocols for the CHOK1SV cell system

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• 作者：M. Celina de la Cruz Edmonds (1)
  Melanie Tellers (1)
  Christine Chan (1)
  Peter Salmon (1)
  David K. Robinson (1)
  Julia Markusen (1)
  
• 关键词：CHO cells ; glutamine synthetase ; MSX ; mammalian cell culture ; cell line stability ; monoclonal antibody production
• 刊名：Molecular Biotechnology
• 出版年：2006
• 出版时间：October 2006
• 年：2006
• 卷：34
• 期：2
• 页码：179-190
• 全文大小：405KB
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• 作者单位：M. Celina de la Cruz Edmonds (1)
  Melanie Tellers (1)
  Christine Chan (1)
  Peter Salmon (1)
  David K. Robinson (1)
  Julia Markusen (1)
  
  1. Merck Research Labs, 07065, Rahway, NJ
  

文摘

To date, the FDA has approved 18 monoclonal antibody (MAb) therapeutic drugs with targets ranging from asthma and rheumatoid arthritis to leukemia. Many of these approved products are produced in Chinese hamster ovary cells (CHO) making CHO a significant and relevant host system. We studied the applicability of CHOK1SV cells as a potential host cell line for MAb production in terms of timelines, achievable titers, transfectant stability, and reproducibility. CHOK1SV, developed by Lonza Biologics, is a suspension, protein-free-adapted CHOK1-derivative utilizing the glutamine synthetase (GS) gene expression system. CHOK1SV expresses the GS enzyme endogenously; thus, positive transfectants were obtained under the dual selection of methionine sulfoximine (MSX) and glutamine-free media. We examined outgrowth efficiencies, specific productivities, and achievable batch titers of three different IgG MAbs transfected into CHOK1SV. Reducing the MSX concentration in the initial selection medium resulted in a decreased incubation time required for transfectant colonies to appear. Specific productivities of “high-producers-ranged between 11 and 49 pg/c/d with batch titers ranging from 105 to 519 mg/L. Transfectant stability and the effects of MSX also were investigated, which indicated that the addition of MSX was necessary to maintain stable MAb production. Cell growth was stable regardless of MSX concentration.