Sensitive real-time on-line estimator for oxygen transfer rates in fermenters
Sensitive real-time on-line estimator for oxygen transfer rates in fermenters
复制标题
发酵罐中氧气传输率的灵敏实时在线估算器
DOI：
10.1016/j.jbiotec.2022.09.005
复制
发表时间：
2022
影响因子：
4.1
通讯作者：
Groff, Richard E.
中科院分区：
文献类型：
--
作者：
Trout, Marshall;Harcum, Sarah W.;Groff, Richard E.
在大型发酵罐中培养的重组大肠杆菌被广泛用于生产质粒和生物制药。控制培养物生长的一种常用方法是预先设定葡萄糖投料，通常是指数投料。预先设定的饲养模式不能自动调整代谢状态的变化，如重组蛋白表达相关的代谢负担或高细胞密度相关的应激。由于培养物耗氧率反映了培养物的代谢状态，有几种方法来估计氧摄取率（OUR）。这些常见的OUR方法应用有限，因为这些方法要么破坏氧气供应，依赖于经验关系，要么无法解释延迟和过滤效应。在这项研究中，开发了一个氧传递速率（OTR）估计器来帮助我们的预测。这种非干扰OTR估计器同时使用溶解氧和废气氧浓度。这种新的OTR估计器捕获了由于发酵罐的物理和化学操作而引起的OTR的微小变化，例如搅拌速度变化，葡萄糖进料速率变化和重组蛋白表达。由于其灵敏度，这种非中断实时OTR估计器可以与饲料控制算法集成，以保持培养物的代谢状态到所需的设定值。
RecombinantEscherichia coligrown in large-scale fermenters are used extensively to produce plasmids and biopharmaceuticals. One method commonly used to control culture growth is predefined glucose feeding, often an exponential feeding profile. Predefined feeding profiles cannot adjust automatically to metabolic state changes, such as the metabolic burden associated with recombinant protein expression or high-cell density associated stresses. As the culture oxygen consumption rates indicates a culture’s metabolic state, there exist several methods to estimate the oxygen uptake rate (OUR). These common OUR methods have limited application since these approaches either disrupt the oxygen supply, rely on empirical relationships, or are unable to account for latency and filtering effects. In this study, an oxygen transfer rate (OTR) estimator was developed to aid OUR prediction. This non-disruptive OTR estimator uses the dissolved oxygen and the off-gas oxygen concentration, in parallel. This new OTR estimator captures small variations in OTR due to physical and chemical manipulations of the fermenter, such as in stir speed variation, glucose feeding rate change, and recombinant protein expression. Due its sensitivity, this non-disruptive real-time OTR estimator could be integrated with feed control algorithms to maintain the metabolic state of a culture to a desired setpoint.