Process Development Engineering, Genentech, Inc., South San Francisco, CA, USA.
Biotechnol Prog. 2010 Jul-Aug;26(4):1200-3. doi: 10.1002/btpr.414.
Disposable technology is being used more each year in the biotechnology industry. Disposable bioreactors allow one to avoid expenses associated with cleaning, assembly and operations, as well as equipment validation. The WAVE bioreactor is well established for Chinese Hamster Ovary (CHO) production, however, it has not yet been thoroughly tested for E. coli production because of the high oxygen demand and temperature maintenance requirements of that platform. The objective of this study is to establish a robust process to generate inoculum for E. coli production fermentations in a WAVE bioreactor. We opted not to evaluate the WAVE system for production cultures because of the high cell densities required in our current E. coli production processes. Instead, the WAVE bioreactor 20/50 system was evaluated at laboratory scale (10-L) to generate inoculum with target optical densities (OD(550)) of 15 within 7-9 h (pre-established target for stainless steel fermentors). The maximum settings for rock rate (40 rpm) and angle (10.5) were used to maximize mass transfer. The gas feed was also supplemented with additional oxygen to meet the high respiratory demand of the culture. The results showed that the growth profiles for the inoculum cultures were similar to those obtained from conventional stainless steel fermentors. These inoculum cultures were subsequently inoculated into 10-L working volume stainless steel fermentors to evaluate the inocula performance of two different production systems during recombinant protein production. The results of these production cultures using WAVE inocula showed that the growth and recombinant protein production was comparable to the control data set. Furthermore, an economic analysis showed that the WAVE system would require less capital investment for installation and operating expenses would be less than traditional stainless steel systems.
一次性技术在生物技术行业中的应用逐年增加。一次性生物反应器可以避免与清洁、组装和操作相关的费用，以及设备验证。WAVE 生物反应器已广泛应用于中国仓鼠卵巢（CHO）生产，但由于该平台对氧气需求和温度维持的要求较高，尚未对大肠杆菌生产进行彻底测试。本研究的目的是建立一个稳健的工艺，以在 WAVE 生物反应器中生成大肠杆菌生产发酵的接种物。我们选择不评估 WAVE 系统用于生产培养物，因为我们当前的大肠杆菌生产过程需要高细胞密度。相反，在实验室规模（10 升）下评估了 WAVE 生物反应器 20/50 系统，以在 7-9 小时内生成目标光密度（OD(550)）为 15 的接种物（不锈钢发酵罐的预先设定目标）。使用最大的搅拌速度（40 rpm）和角度（10.5）来最大化质量传递。还补充了气体进料以额外的氧气来满足培养物的高呼吸需求。结果表明，接种物培养物的生长曲线与从传统不锈钢发酵罐获得的生长曲线相似。这些接种物培养物随后被接种到 10 升工作体积的不锈钢发酵罐中，以评估两种不同生产系统在重组蛋白生产过程中的接种物性能。使用 WAVE 接种物的这些生产培养物的结果表明，生长和重组蛋白生产与对照数据集相当。此外，经济分析表明，WAVE 系统的安装资本投资较少，运营费用也将低于传统的不锈钢系统。
