Development of monoclonal antibody cell culture processes and antibody characterisation

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dc.contributor.advisorAllen, Evin
dc.contributor.advisorZhdanov, Alexander
dc.contributor.authorGogia, Shreyaen
dc.date.accessioned2026-05-21T15:21:46Z
dc.date.available2026-05-21T15:21:46Z
dc.date.issued2025-11-04en
dc.date.submitted2025-11-04
dc.description.abstractMonoclonal 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.en
dc.description.statusNot peer revieweden
dc.description.versionAccepted Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationGogia, S. 2025. Development of monoclonal antibody cell culture processes and antibody characterisation. MRes Thesis, University College Cork.
dc.identifier.endpage144
dc.identifier.urihttps://hdl.handle.net/10468/18814
dc.language.isoen
dc.publisherUniversity College Corken
dc.rights© 2025, Shreya Gogia.
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectMonoclonal antibodies
dc.subjectUpstream and downstream processing
dc.subjectAnti-IL-8
dc.subjectCHO-DP12 cell line
dc.titleDevelopment of monoclonal antibody cell culture processes and antibody characterisation
dc.typeMasters thesis (Research)en
dc.type.qualificationlevelMasters
dc.type.qualificationnameMRes - Master of Research
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