A predictive high‐throughput scale‐down model of monoclonal antibody production in CHO cells

Abstract

Multi-factorial experimentation is essential in understanding the link between mammalian cell culture conditions and the glycoprotein product of any biomanufacturing process. This understanding is increasingly demanded as bioprocess development is influenced by the Quality by Design paradigm. We have developed a system that allows hundreds of micro-bioreactors to be run in parallel under controlled conditions, enabling factorial experiments of much larger scope than is possible with traditional systems. A high-throughput analytics workflow was also developed using commercially available instruments to obtain product quality information for each cell culture condition. The micro-bioreactor system was tested by executing a factorial experiment varying four process parameters: pH, dissolved oxygen, feed supplement rate, and reduced glutathione level. A total of 180 micro-bioreactors were run for 2 weeks during this DOE experiment to assess this scaled down micro-bioreactor system as a high-throughput tool for process development. Online measurements of pH, dissolved oxygen, and optical density were complemented by offline measurements of glucose, viability, titer, and product quality. Model accuracy was assessed by regressing the micro-bioreactor results with those obtained in conventional 3 L bioreactors. Excellent agreement was observed between the micro-bioreactor and the bench-top bioreactor. The micro-bioreactor results were further analyzed to link parameter manipulations to process outcomes via leverage plots, and to examine the interactions between process parameters. The results show that feed supplement rate has a significant effect (P < 0.05) on all performance metrics with higher feed rates resulting in greater cell mass and product titer. Culture pH impacted terminal integrated viable cell concentration, titer and intact immunoglobulin G titer, with better results obtained at the lower pH set point. The results demonstrate that a micro-scale system can be an excellent model of larger scale systems, while providing data sets broader and deeper than are available by traditional methods.

Cell Culture Processes in Monoclonal Antibody Production

“…77,78 This regulatory requirement reflects the recognition to tens of milliliters of working volumes, are also being evaluated [86][87][88] for robustness, reliability and scalability. It is conceivable that one or more of these systems could be used more widely for automated screening of a large number of culture conditions.…”

Cell Culture Processes for Biopharmaceutical Manufacturing

“…Simcell was among the first microbioreactor to be commercially available with culture volumes of less than a millilitre 62 . Later microbioreactor platforms, several of which are now available commercially, were developed to culture larger culture volumes in the millilitres range, which may also enable expression of greater amounts of product for better characterization 63 .…”

Fermentative Biohydrogen Modelling and Optimization Research in Light of Miniaturized Parallel Bioreactors

“…therefore, microbioreactor technology has been used to address these challenges in order to reduce experimentation costs and speed up the research output. Industries have often employed simplified systems such as microtiter plates, shake flasks, test-tubes and spinner flasks for multi-factorial experimentation which offer ideal strategy to investigate the complex relationships between culture conditions and process outcomes (5,36). Several authors have highlighted the need for miniaturized parallel bioreactors that monitors and controls the physicochemical parameters for high throughput experimentation (6,7,22,24,42,59,61).…”