Very high density of Chinese hamster ovary cells in perfusion by alternating tangential flow or tangential flow filtration in WAVE Bioreactor™-part II: Applications for antibody production and cryopreservation.
Very high density of Chinese hamster ovary cells in perfusion by alternating tangential flow or tangential flow filtration in WAVE Bioreactor™-part II: Applications for antibody production and cryopreservation.
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DOI：
10.1002/btpr.1703
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发表时间：
2013-05
影响因子：
2.9
通讯作者：
Chotteau, Veronique
中科院分区：
文献类型：
--
作者：
Clincke, Marie-Francoise;Molleryd, Carin;Samani, Puneeth K.;Lindskog, Eva;Faldt, Eric;Walsh, Kieron;Chotteau, Veronique
关键词：
在一次性WAVE Bioreactor™中开发了生产单克隆抗体（MAb）的中国仓鼠卵巢（CHO）细胞的高细胞密度灌注方法，使用外部中空纤维（HF）过滤器作为细胞分离装置。比较了切向流过滤（TFF）和交替切向流（ATF）系统，并研究了高细胞密度灌注的工艺应用：单克隆抗体生产和冷冻保存。通过微滤（MF）或超滤（UF）与ATF或TFF的灌注操作和通过分批补料进行了比较。使用UF TFF或UF ATF获得高于108个细胞/mL的细胞密度。通过ATF或TFF、MF或UF灌注，细胞产生相当量的MAb。使用ATF或TFF的MF部分保留了MAb，但使用TFF时更严重。因此，当在每日采血中从生物反应器中丢弃细胞肉汤时，MAb丢失。灌注中细胞密度高达1.3 × 108个细胞/mL时的MAb细胞比生产率相当，补料分批中的MAb细胞比生产率相当或更低。12天后，与补料分批相比，使用ATF或TFF与MF或UF的灌注收获了6倍多的MAb，并且在108个细胞/mL密度的1个月灌注中收获了28倍多的MAb。以高达2.75 L/min的再循环速率泵送不会损坏具有HF短路的本TFF设置的细胞。使用来自108个细胞/mL密度灌注运行的细胞，以0.5 × 108和108个细胞/mL进行细胞冷冻保存。细胞复苏非常成功，表明该系统是一种可靠的细胞库生产工艺。© 2013美国化学工程师学会生物技术。程序：29：768-777，2013
A high cell density perfusion process of monoclonal antibody (MAb) producing Chinese hamster ovary (CHO) cells was developed in disposable WAVE Bioreactor™ using external hollow fiber (HF) filter as cell separation device. Tangential flow filtration (TFF) and alternating tangential flow (ATF) systems were compared and process applications of high cell density perfusion were studied here: MAb production and cryopreservation. Operations by perfusion using microfiltration (MF) or ultrafiltration (UF) with ATF or TFF and by fed-batch were compared. Cell densities higher than 108 cells/mL were obtained using UF TFF or UF ATF. The cells produced comparable amounts of MAb in perfusion by ATF or TFF, MF or UF. MAbs were partially retained by the MF using ATF or TFF but more severely using TFF. Consequently, MAbs were lost when cell broth was discarded from the bioreactor in the daily bleeds. The MAb cell-specific productivity was comparable at cell densities up to 1.3 × 108 cells/mL in perfusion and was comparable or lower in fed-batch. After 12 days, six times more MAbs were harvested using perfusion by ATF or TFF with MF or UF, compared to fed-batch and 28× more in a 1-month perfusion at 108 cells/mL density. Pumping at a recirculation rate up to 2.75 L/min did not damage the cells with the present TFF settings with HF short circuited. Cell cryopreservation at 0.5 × 108 and 108 cells/mL was performed using cells from a perfusion run at 108 cells/mL density. Cell resuscitation was very successful, showing that this system was a reliable process for cell bank manufacturing. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:768–777, 2013
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