Early Stage Cell Culture, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
Biotechnol Prog. 2011 Sep-Oct;27(5):1397-406. doi: 10.1002/btpr.659. Epub 2011 Jul 11.
The biopharmaceutical industry is increasing its use of the WAVE Bioreactor for culturing cells. Although this disposable bioreactor can be equipped to provide real-time pH and dissolved oxygen (DO) monitoring and control, our goal was to develop a process for culturing CHO cells in this system without relying on pH and DO feedback controls. After identifying challenges in culturing cells without controlling for pH and DO in the WAVE Bioreactor, we characterized O(2) and CO(2) transfer in the system. From these cell-free studies, we identified rock rate and rock angle as key parameters affecting O(2) transfer. We also identified the concentration of CO(2) in the incoming gas and the rate of gas flow into the headspace as key parameters affecting CO(2) transfer--and therefore pH--in the disposable culture chamber. Using a full-factorial design to evaluate the rock rate, rock angle, and gas flow rate defined for this WAVE Bioreactor process, we found comparable cell growth and pH profiles in the ranges tested for these three parameters in two CHO cell lines. This process supported cell growth, and maintained pH and DO within our desired range--pH 6.8-7.2 and DO exceeding 20% of air saturation--for six CHO cell lines, and it also demonstrated comparable cell growth and viability with the stirred-tank bioreactor process with online pH and DO control. By eliminating the use of pH and DO probes, this process provides a simple and more cost-effective method for culturing cells in the WAVE Bioreactor.
生物制药行业越来越多地使用 WAVE 生物反应器来培养细胞。虽然这种一次性生物反应器可以配备实时 pH 值和溶解氧 (DO) 监测和控制功能，但我们的目标是开发一种在不依赖 pH 值和 DO 反馈控制的情况下在该系统中培养 CHO 细胞的工艺。在确定了在不控制 WAVE 生物反应器中 pH 值和 DO 的情况下培养细胞的挑战后，我们对该系统中的 O(2) 和 CO(2) 转移进行了特征描述。通过这些无细胞研究，我们确定了摇床速度和摇床角度是影响 O(2)转移的关键参数。我们还确定了进入气体中的 CO(2)浓度和进入头部空间的气体流速是影响 CO(2)转移（从而影响 pH 值）的关键参数在一次性培养室中。使用全因子设计来评估此 WAVE 生物反应器工艺中定义的摇床速度、摇床角度和气体流速，我们发现这三个参数在两个 CHO 细胞系的测试范围内具有可比的细胞生长和 pH 曲线。该工艺支持细胞生长，并将 pH 值和 DO 值维持在我们所需的范围内——pH 值为 6.8-7.2，DO 值超过空气饱和值的 20%——用于六种 CHO 细胞系，并且它还与具有在线 pH 值和 DO 值控制的搅拌罐生物反应器工艺具有可比的细胞生长和活力。通过消除 pH 值和 DO 探头的使用，该工艺为在 WAVE 生物反应器中培养细胞提供了一种简单且更具成本效益的方法。
