Defining process design space for monoclonal antibody cell culture

Abstract

The concept of design space has been taking root as a foundation of in-process control strategies for biopharmaceutical manufacturing processes. During mapping of the process design space, the multidimensional combination of operational variables is studied to quantify the impact on process performance in terms of productivity and product quality. An efficient methodology to map the design space for a monoclonal antibody cell culture process is described. A failure modes and effects analysis (FMEA) was used as the basis for the process characterization exercise. This was followed by an integrated study of the inoculum stage of the process which includes progressive shake flask and seed bioreactor steps. The operating conditions for the seed bioreactor were studied in an integrated fashion with the production bioreactor using a two stage design of experiments (DOE) methodology to enable optimization of operating conditions. A two level Resolution IV design was followed by a central composite design (CCD). These experiments enabled identification of the edge of failure and classification of the operational parameters as non-key, key or critical. In addition, the models generated from the data provide further insight into balancing productivity of the cell culture process with product quality considerations. Finally, process and product-related impurity clearance was evaluated by studies linking the upstream process with downstream purification. Production bioreactor parameters that directly influence antibody charge variants and glycosylation in CHO systems were identified.

7.3 Process Characterization for Upstream and Downstream Process Development

“…Range 1, which is within the operating limits, may be too narrow to give a detectable signal over noise ratio, and the parameter may not have a statistically significant effect on the response(s). For example Abu-Absi et al [46] found from screening studies that temperature did not affect Brought to you by | New York University Bobst Library Technical Services Authenticated Download Date | 7/6/15 12:03 PM N-glycosylation. However, when they widened the temperature range tested, a quadratic model was built and temperature was identified as a critical process parameter (CPP).…”

Tailoring recombinant protein quality by rational media design

“…The temperature shift led to an overall increase, amounting to 2.3-5.6% of HM. Mild hypothermia has been shown to impact galactosylation depending on media composition, cell line and protein type [277][278][279] . As expected based upon biological knowledge, the media supplements weakly influenced the abundance of afucosylated species.…”

Establishment of a Design Space for Biopharmaceutical Purification Processes Using DoE

“…DoE has been used in many fields of application besides protein purification [8,9], such as development of culture media either for prokaryotic or eukaryotic cells [10], recombinant protein and virus productivity optimization [11,12].…”