Programa de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Coyoacán, Ciudad De México, México.
Bioengineered. 2020 Dec;11(1):463-471. doi: 10.1080/21655979.2020.1744266.
A careful selection of culture mediums and feeds has become necessary to maximize yields of recombinant proteins during bioprocesses of mammalian cells. Supplements contain a variety of concentrate nutrients, and their beneficial effects vary according to recombinant cell lines. In this study, the effects of PowerFeed A on growth kinetics, productivity and cellular metabolism were evaluated for two Chinese hamster ovary cell lines producing a monoclonal antibody in a batch culture. Supplemented cultures increased integral viable cell density of CRL-12444 and CRL-12445 cells by 2.4 and 1.6 times through extension of culture time at which viability was above 90% in 72 and 36 h, respectively, and increment of maximal cell concentration in 3.25 × 10 cells/ml (69%) for CRL-12445 cells. Product titer augmented 1.9 and 2.5 times for CRL-12444 and CRL-12445 cells, respectively, without changes in growth rate and specific productivity. Feed supplementation also stimulated full consumption of glucose and free glutamine and reduced 10 times lactate accumulation, while ammonium, sodium and potassium remained at similar concentrations at the end of the culture. About 44% of calcium, mainly provided by feed, was consumed by both cell lines. Maximization of cellular growth, viability and protein titer through feeding encourages extending its use to other cell lines and exploring novel combinations with other basal mediums or feeds. A thorough investigation of its impact on protein quality and the molecular mechanisms behind these effects will allow designing effective feeds and strategies to rationally optimize protein production in the biomanufacturing industry.
在哺乳动物细胞的生物工艺过程中，为了最大限度地提高重组蛋白的产量，必须仔细选择培养基和饲料。补充剂含有各种浓缩营养素，其有益效果因重组细胞系而异。在这项研究中，评估了 PowerFeed A 对生产单克隆抗体的两种中国仓鼠卵巢细胞系在批式培养中的生长动力学、生产力和细胞代谢的影响。补充培养通过延长培养时间，使 CRL-12444 和 CRL-12445 细胞的存活率保持在 90%以上分别达到 72 小时和 36 小时，使最大细胞浓度分别增加 3.25×10 细胞/ml（69%），使 CRL-12445 细胞的整体活细胞密度分别增加了 2.4 倍和 1.6 倍。CRL-12444 和 CRL-12445 细胞的产物滴度分别增加了 1.9 倍和 2.5 倍，而生长速率和比生产率没有变化。补料还刺激了葡萄糖和游离谷氨酰胺的完全消耗，将乳酸积累减少了 10 倍，而氨、钠和钾在培养结束时仍保持在相似的浓度。两条细胞系消耗的钙约有 44%，主要来自饲料。通过补料实现细胞生长、活力和蛋白滴度的最大化，鼓励将其应用于其他细胞系，并探索与其他基础培养基或饲料的新组合。深入研究其对蛋白质质量的影响以及这些影响背后的分子机制，将有助于设计有效的饲料和策略，从而在生物制造行业中合理优化蛋白质生产。
