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首页> 外文会议>Conference on single-use technologies II: bridging polymer science to biotechnology applications >PERFUSION MICROBIOREACTOR WITH INTEGRATED CELL RETENTION DEVICE
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PERFUSION MICROBIOREACTOR WITH INTEGRATED CELL RETENTION DEVICE

机译：集成细胞保留装置的灌注微生物反应器

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Intensified perfusion processes are a key component in Integrated Continuous Biomanufacturing that still faces many challenges. For example, delivering the right amount of nutrients for extremely high cell densities at economically feasible perfusion rates need the right cell culture media and process development. To this end, data sets are generated using high-throughput small-scale models. In the absence of a cell retention device that can work with those small working volumes, strategies that mimic perfusion are being used. For example, a common technique is to use either high density batch or simulation of perfusion in spin tubes or ambr®15 using discrete media exchanges. However, the applicability of the data generated in these small-scale models is limited and process development must be supplemented with perfusion bioreactors at benchtop scale. The key point that these small-scale models miss to accurately predict perfusion processes is a cell retention device that enables continuous media exchange while retaining cells in the bioreactor. In this work we will present the evaluation of a perfusion microbioreactor system. This 2mL perfusion microbioreactor has all the requirements to accurately control DO, pH and temperature; it is equipped with a filtration-based cell retention device and optical density sensors that enable the performance of continuous perfusion with automated cell bleed (Figure 1 ). We will show cell performance in the perfusion microbioreactor system at 2wd using a CHOZN®-GS producing a fusion protein (n=3) and a CHO-S producing an IgG (n=2) in steady state with a target viable cell density of 50×10~6vc/mL and dynamic perfusion. We will then compare cell growth, metabolites and production to steady state and dynamic 3L perfusion bioreactors and we will review process modifications made during the evaluation, including gas and mixing strategies. Lastly, we will present two case studies using the perfusion microbioreactor system: 1) Evaluation of cell performance in three different media. 2) Determination of minimum CSPR.
机译：强化的灌注过程是集成连续生物制造中的关键组成部分，仍然面临许多挑战。例如，以经济上可行的灌注速率为极高的细胞密度提供适量的营养，需要正确的细胞培养基和工艺开发。为此，使用高通量小规模模型来生成数据集。在没有可以与那些小工作量一起使用的细胞保留装置的情况下，正在使用模仿灌注的策略。例如，一种常见的技术是使用高密度批次或使用离散介质交换在旋转管或ambr®15中模拟灌注。但是，在这些小规模模型中生成的数据的适用性受到限制，并且必须在台式规模上使用灌注生物反应器来补充过程开发。这些小规模模型无法准确预测灌注过程的关键点是细胞保留设备，该设备能够在将细胞保留在生物反应器中的同时进行连续的培养基交换。在这项工作中，我们将介绍灌注微生物反应器系统的评估。这款2mL灌注微生物反应器具有精确控制DO，pH和温度的所有要求。它配备了基于过滤的细胞滞留装置和光密度传感器，可通过自动细胞流血实现连续灌注（图1）。我们将在2wd使用稳定的CHOZN®-GS产生融合蛋白（n = 3）和CHO-S产生IgG（n = 2）的CHOZN®-GS展示其在灌注微生物反应器系统中的细胞性能，目标活细胞密度为50×10〜6vc / mL，动态灌注。然后，我们将细胞生长，代谢产物和产量与稳态和动态3L灌注生物反应器进行比较，并评估在评估过程中进行的工艺修改，包括气体和混合策略。最后，我们将介绍使用灌注微生物反应器系统的两个案例研究：1）评估三种不同培养基中的细胞性能。 2）确定最低CSPR。

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