Scale‐down of CHO cell cultivation from shake flasks based on oxygen mass transfer allows application of parallelized, non‐invasive, and time‐resolved monitoring of the oxygen transfer rate in 48‐well microtiter plates

Abstract

Cultivating Chinese hamster ovary (CHO) cells in microtiter plates (MTPs) with time‐resolved monitoring of the oxygen transfer rate (OTR) is highly desirable to provide process insights at increased throughput. However, monitoring of the OTR in MTPs has not been demonstrated for CHO cells, yet. Hence, a CHO cultivation process was transferred from shake flasks to MTPs to enable monitoring of the OTR in each individual well of a 48‐well MTP. For this, the cultivation of an industrially relevant, antibody‐producing cell line was transferred from shake flask to MTP based on the volumetric oxygen mass transfer coefficient (kLa). Culture behavior was well comparable (deviation of the final IgG titer less than 10%). Monitoring of the OTR in 48‐well MTPs was then used to derive the cytotoxicity of dimethyl sulfoxide (DMSO) based on a dose–response curve in a single experiment using a second CHO cell line. Logistic fitting of the dose–response curve determined after 100 h was used to determine the DMSO concentration that resulted in a cytotoxicity of 50% (IC50). A DMSO concentration of 2.70% ± 0.25% was determined, which agrees with the IC50 previously determined in shake flasks (2.39% ± 0.1%). Non‐invasive, parallelized, and time‐resolved monitoring of the OTR of CHO cells in MTPs was demonstrated and offers excellent potential to speed up process development and assess cytotoxicity.

Recombinant human fibroblast growth factor 7 obtained from stable Chinese hamster ovary cells enhances wound healing

“…[25] To address the limitations of HEK293 cells, culture conditions can be optimized, or alternative cell lines such as CHO can be used. [26,27] CHO cells have a lower metabolic rate and are capable of producing high-quality glycosylated proteins, making them promising platforms for biopharmaceutical manufacturing. [27] In fact, CHO cells are already used to produce FDA and EMA approved biopharmaceuticals, highlighting the promising applicability of CHO in the pharmaceutical industry [28,33] and suggesting its potential for FGF7 production.…”

“…[26,27] CHO cells have a lower metabolic rate and are capable of producing high-quality glycosylated proteins, making them promising platforms for biopharmaceutical manufacturing. [27] In fact, CHO cells are already used to produce FDA and EMA approved biopharmaceuticals, highlighting the promising applicability of CHO in the pharmaceutical industry [28,33] and suggesting its potential for FGF7 production. However, to the best of our knowledge, no previous studies have reported on the application of CHO cells for recombinant FGF7 production.…”