Department of Biotechnology, Institute of Bioprocess Science and Engineering, University of Natural Resources and Life Sciences, Vienna, Austria.
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, Netherlands.
Biotechnol Bioeng. 2021 Sep;118(9):3382-3394. doi: 10.1002/bit.27738. Epub 2021 Mar 25.
Chemically defined media are reconstituted batchwise and stored in hold tanks until use. To avoid large hold tanks and batchwise production of media, we developed continuous on-demand reconstitutions directly from solids consisting of a hopper and a screw conveyor capable of feeding dry powdered media with the required precision ±5% at low dosing rates of 0.171 g min . A commercially available dry powdered cell culture medium was continuously fed over a duration of 12 h into a mixer which was connected to a UV-cell for monitoring and the media were compared to a batchwise production. A comparable amino acid, carbohydrate, and osmolality profile to a batchwise reconstitution could be obtained. Cell cultivation showed comparable performance of batch and continuous reconstitution for two CHO cell lines producing the antibodies adalimumab and trastuzumab on a small and benchtop scale. In-depth analysis of the produced antibodies showed the same glycosylation pattern, other posttranslational profiles such as methionine oxidation and deamidation compared to batchwise reconstitution. Therefore, we conclude a continuous reconstitution of the medium results in the same quality of the product. A continuous on-demand media reconstitution will impact the supply chain and significantly reduce the floor space necessary for preparation and storage.
化学定义的培养基是分批复配的，并储存在储罐中，直到使用。为了避免使用大型储罐和分批生产培养基，我们开发了一种连续按需从固体中直接复配的方法，固体由料斗和螺旋输送机组成，能够以低投加率（0.171g/min）以±5%的精度输送干粉状培养基。一种市售的干粉细胞培养基在 12 小时内连续进料到混合器中，混合器连接到 UV 细胞以进行监测，并将培养基与分批生产进行比较。可以获得与分批复配相当的氨基酸、碳水化合物和渗透压轮廓。细胞培养表明，两种 CHO 细胞系在小规模和台式规模上生产阿达木单抗和曲妥珠单抗时，分批和连续复配的性能相当。对产生的抗体进行深入分析表明，与分批复配相比，其糖基化模式相同，其他翻译后谱如蛋氨酸氧化和脱酰胺作用相同。因此，我们得出结论，连续复配培养基会导致产品质量相同。连续按需的培养基复配将影响供应链，并显著减少准备和储存所需的占地面积。
