Development of monoclonal antibody cell culture processes and antibody characterisation
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Date
2025-11-04
Authors
Gogia, Shreya
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Publisher
University College Cork
Published Version
Abstract
Monoclonal antibodies (mAbs) have transformed modern therapeutics due to their high specificity and efficacy for a wide range of diseases. This research project aimed to establish an end-to-end laboratory-scale monoclonal antibody production process, including upstream cell culture, downstream purification and analytical characterisation. A CHO-DP12 cell line engineered for anti-IL-8 mAb expression and a cNIST mAb-producing CHO cell line were employed to investigate key cell culture process parameters.
Upstream studies examined the effects of media formulations and seeding density studies on antibody expression. While serum-supplemented media supported the growth of CHO-DP12 cells, the serum-derived impurities and inconsistent antibody expression limited the production of anti-IL-8 mAbs. In contrast, CHO-NIST cells cultured in manufacturer-recommended, serum-free media successfully produced detectable antibodies.
Analytical characterisation was performed using SDS-PAGE, Western Blotting and SEC-HPLC to assess protein concentration, purity and structural integrity. Stability and forced degradation studies were further conducted to assess the impact of stressful conditions on antibody integrity.
Preliminary cell-based assays were explored using HL-60 and HCT-116 cell lines to study IL-8 signalling and anti-IL-8 mAb activity. These assays generated variable and inconclusive results that highlighted the need for further work using alternative mechanism-specific functional assays.
Overall, while limited in final results, the work advanced the current state of mAb production within the research group and provides a roadmap towards anti-IL-8 mAb production and potency assessment.
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Keywords
Monoclonal antibodies , Upstream and downstream processing , Anti-IL-8 , CHO-DP12 cell line
Citation
Gogia, S. 2025. Development of monoclonal antibody cell culture processes and antibody characterisation. MRes Thesis, University College Cork.