In order to study the impact of zinc and copper on the titer levels of mAb and recombinant protein in CHO cells, the IgG-expressing (DP12) and EPO-expressing (SK15) cell lines were cultured in chemically defined media with increasing concentrations of either metal. Supplementation with 25 mg/l in CDM media resulted in a significant increase in EPO (1.7-fold) and IgG (2.6-fold) titers compared to control (no added zinc). Titers at this Zn concentration in CDM containing the insulin replacing agent aurintricarboxylic acid (ATA) (CDM + A) showed a 1.8-fold (EPO) and 1.2-fold (IgG) titers increase compared to control. ATA appeared to also reduce the specific productivity (Qp) enhancement induced by Zn-25, with up to 4.9-fold (DP12) and 1.9-fold (SK15) Qp increase in CDM compared to the 1.6-fold (DP12) and 1.5-fold (SK15) Qp increase observed in CDM + A. A 31% reduced Viable Cell Density (VCD) in DP12 was observed in both Zn-supplemented media (3 × 106 cells/ml vs 4.2 × 106 cells/ml, day 5), whereas SK15 Zn-25 cultures displayed a 24% lower peak only in CDM + A (2.2 × 106 cells/ml vs 3.2 × 106 cells/ml, day 5). Supplementation with copper at 13.7-20 mg/l resulted in less significant cell line/product-type dependent effects on titer, VCD and Viability. Analysis of the energetic phenotype of both cell lines in 25 mg/l Zn-supplemented CDM media revealed a twofold increase in the oxygen consumption rate (OCR) compared to non-supplemented cells. Together, these data suggest that high zinc supplementation may induce an increase in oxidative respiration metabolism that results in increased Qp and titers in suspension CHO cultures.

中文翻译：

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补充锌可增加重组 CHO 细胞的蛋白质效价。

为了研究锌和铜对 CHO 细胞中 mAb 和重组蛋白滴度水平的影响，将表达 IgG (DP12) 和表达 EPO (SK15) 的细胞系在化学成分确定的培养基中培养，其中任一浓度递增。金属。与对照（未添加锌）相比，在 CDM 培养基中添加 25 mg/l 导致 EPO（1.7 倍）和 IgG（2.6 倍）滴度显着增加。与对照相比，含有胰岛素替代剂金精三羧酸 (ATA) (CDM + A) 的 CDM 中在此 Zn 浓度下的滴度显示出 1.8 倍 (EPO) 和 1.2 倍 (IgG) 滴度增加。ATA 似乎还降低了 Zn-25 引起的比生产力 (Qp) 增强，与 1.6 倍 (DP12) 和 1.5 倍相比，CDM 中的 Qp 增加高达 4.9 倍 (DP12) 和 1.9 倍 (SK15)。在 CDM + A 中观察到 Qp 增加 (SK15) 倍。在两种补充 Zn 的培养基中观察到 DP12 中的活细胞密度 (VCD) 降低了 31%（3 × 106 个细胞/ml 与 4.2 × 106 个细胞/ml，第 5 天） ，而 SK15 Zn-25 培养物仅在 CDM + A 中显示出 24% 的较低峰值（2.2 × 106 细胞/ml 与 3.2 × 106 细胞/ml，第 5 天）。补充 13.7-20 mg/l 的铜对滴度、VCD 和活力的细胞系/产品类型依赖性影响不太显着。对 25 mg/l 补充 Zn 的 CDM 培养基中两种细胞系的能量表型分析表明，与未补充的细胞相比，耗氧率 (OCR) 增加了一倍。总之，这些数据表明，高锌补充可能会诱导氧化呼吸代谢增加，从而导致悬浮 CHO 培养物中 Qp 和滴度增加。