Development of monoclonal antibody cell culture processes and antibody characterisation

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.

Grantor dc:publisher

University College Cork

Year dc:date.issued

2025

Author dc:creator

• Gogia, Shreya

Advisors dc:contributor.advisor

• Allen, Evin
• Zhdanov, Alexander

• Monoclonal antibodies
• Upstream and downstream processing
• Anti-IL-8
• CHO-DP12 cell line

Statement dc:rights

• © 2025, Shreya Gogia.

Licence dc:rights.uri

• https://creativecommons.org/licenses/by/4.0/

Language dc:language.iso

en

Handle dc:identifier.uri

https://hdl.handle.net/10468/18814

OAI identifier oai:identifier

oai:cora.ucc.ie:10468/18814