Advances and Drawbacks of the Adaptation to Serum-Free Culture of CHO-K1 Cells for Monoclonal Antibody Production

Abstract

Currently, mammalian cell technology has become the focus of biopharmaceutical production, with strict regulatory scrutiny of the techniques employed. Major concerns about the presence of animal-derived components in the culture media led to the development of serum-free (SF) culture processes. However, cell adaptation to SF conditions is still a major challenge and limiting step of process development. Thus, this study aims to assess the impact of SF adaptation on monoclonal antibody (mAb) production, identify the most critical steps of cell adaptation to the SF EX-CELL medium, and create basic process guidelines. The success of SF adaptation was dependent on critical steps that included accentuated cell sensitivity to common culture procedures (centrifugation, trypsinization), initial cell concentration, time given at each step of serum reduction, and, most importantly, medium supplements used to support adaptation. Indeed, only one of the five supplement combinations assessed (rhinsulin, ammonium metavanadate, nickel chloride, and stannous chloride) succeeded for the Chinese hamster ovary-K1 cell line used. This work also revealed that the chemically defined EX-CELL medium benefits mAb production in comparison with the general purpose Dulbecco's Modified Eagle's Medium, but the complete removal of serum attenuates these positive effects.

Adaptation of CHO cells in serum‐free conditions for erythropoietin production: Application of EVOP technique for process optimization

“…1). This result suggests that cells need a substantial time during the early stage of the adaptation process to achieve optimal growth [17]. Overall, adequate growth and viability of rCHO cells in serum-free conditions were obtained after an adaptation process of 130 days, which is in accordance with the results of previous studies on the adaptation of CHO cells producing recombinant antibodies [18,19].…”

Monitoring changes in proteome during stepwise adaptation of a MDCK cell line from adherence to growth in suspension

“…Regarding product safety and batch-to-batch reproducibility, however, adaptation to SFM and finally to suspension growth in CDM is preferred. So far, only few cell lines are available where these challenges have been addressed successfully and adaptation of cell lines is still a difficult task [43].…”

Mucin‐coating technologies for protection and reduced aggregation of cellular production systems

“…However, the ability of our coatings to protect cells and strongly resist clumping could also benefit current biomanufacturing platforms, like CHO cells, which can still aggregate under nonideal reactor conditions or in nonoptimal media formulations. As biomanufacturing looks beyond CHO systems for next‐generation production platforms that mitigate the risk of nonhuman glycoconjugates and other antigenic epitopes, adaptation to growth in suspension remains a significant and time‐consuming challenge for human, primate, and many other mammalian cell lines (do Amaral et al, ; Rodrigues et al, ). By promoting cell viability and minimizing aggregation, our coatings could help overcome some of the significant barriers to suspension adaptation.…”