{"id":{"repo_id":"cork","oai_identifier":"oai:cora.ucc.ie:10468/18814"},"canonical_url":"https://search.dev.ndltd.org/etd/cork/oai:cora.ucc.ie:10468/18814","repository":{"repo_id":"cork","name":"University College Cork","base_url":"https://cora.ucc.ie/server/oai/request"},"display":{"title":"Development of monoclonal antibody cell culture processes and antibody characterisation","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.","abstract_html":"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.","abstract_has_math":false,"creators":["Gogia, Shreya"],"institution":"University College Cork","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Allen, Evin","Zhdanov, Alexander"],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-11-04","date_published":"2025-11-04","updated_at":"2026-07-24T01:46:55Z","subjects":["Monoclonal antibodies","Upstream and downstream processing","Anti-IL-8","CHO-DP12 cell line"],"languages":["en"],"rights":["© 2025, Shreya Gogia."],"rights_urls":["https://creativecommons.org/licenses/by/4.0/"],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/10468/18814","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Allen, Evin","Zhdanov, Alexander"]},{"key":"dc:creator","label":"Author","values":["Gogia, Shreya"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2026-05-21T15:21:46Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2026-05-21T15:21:46Z"]},{"key":"dc:date.issued","label":"Date","values":["2025-11-04"]},{"key":"dc:publisher","label":"Institution","values":["University College Cork"]},{"key":"dc:type","label":"Dc Type","values":["Masters thesis (Research)"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["Masters"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["MRes - Master of Research"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Monoclonal antibodies","Upstream and downstream processing","Anti-IL-8","CHO-DP12 cell line"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["© 2025, Shreya Gogia."]},{"key":"dc:rights.uri","label":"Rights URI","values":["https://creativecommons.org/licenses/by/4.0/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/10468/18814"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["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."]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Development of monoclonal antibody cell culture processes and antibody characterisation"]}]}],"canonical_facts":{"dc:contributor.advisor":["Allen, Evin","Zhdanov, Alexander"],"dc:creator":["Gogia, Shreya"],"dc:date.accessioned":["2026-05-21T15:21:46Z"],"dc:date.available":["2026-05-21T15:21:46Z"],"dc:date.issued":["2025-11-04"],"dc:description.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."],"dc:format.mimetype":["application/pdf"],"dc:identifier.uri":["https://hdl.handle.net/10468/18814"],"dc:language.iso":["en"],"dc:publisher":["University College Cork"],"dc:rights":["© 2025, Shreya Gogia."],"dc:rights.uri":["https://creativecommons.org/licenses/by/4.0/"],"dc:subject":["Monoclonal antibodies","Upstream and downstream processing","Anti-IL-8","CHO-DP12 cell line"],"dc:title":["Development of monoclonal antibody cell culture processes and antibody characterisation"],"dc:type":["Masters thesis (Research)"],"dc:type.qualificationlevel":["Masters"],"dc:type.qualificationname":["MRes - Master of Research"]},"updated_at":"2026-07-24T01:46:55Z"}