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首页> 外文期刊>Biotechnology Progress >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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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

机译：通过交替在WAVE生物反应器中切向流或切向流过滤在灌流中非常高密度的中国仓鼠卵巢细胞-第II部分：抗体生产和冷冻保存的应用

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摘要

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 10`8 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 x 10~8 cellslmL 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 28X more in a 1-month perfusion at 10~8 cells/ mL density. Pumping at a recirculation rate up to 2.75 Llmin did not damage the cells with the present TFF settings with HF short circuited. Cell cryopreservation at 0.5 X 10s and 10s cellslmL was performed using cells from a perfusion run at lCr cellslmL density. Cell resuscitation was very successful, showing that this system was a reliable process for cell bank manufacturing.
机译：在一次性WAVE Bioreactor中开发了一种产生单克隆抗体（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，并在此研究了高细胞密度灌注的过程应用：单抗的生产和冷冻保存。比较了通过使用ATF或TFF的微滤（MF）或超滤（UF）灌流以及分批补料进行的操作。使用UF TFF或UF ATF获得的细胞密度高于10-8个细胞/ mL。在通过ATF或TFF，MF或UF进行灌注时，细胞产生了相当数量的MAb。使用ATF或TFF，MF会部分保留单克隆抗体，但使用TFF会更严重。因此，当每天流血从生物反应器中丢弃细胞肉汤时，MAb就会丢失。在高达1.3 x 10〜8细胞/ mL的细胞密度下，MAb细胞的特异性生产力在灌注中可比，在补料分批中可比或更低。 12天后，使用ATF或TFF与MF或UF进行灌注，收获的单克隆抗体多于补料分批，在1个月的灌注中，以10〜8个细胞/ mL的密度收获的单克隆抗体多于28倍。以当前的TFF设置，HF短路，以高达2.75 Llmin的再循环速率泵送不会损坏细胞。使用来自以1Cr细胞1mL密度的灌注运行的细胞在0.5×10s和10s细胞1mL进行细胞冷冻保存。细胞复苏非常成功，表明该系统是用于细胞库制造的可靠过程。

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