Development of an end-to-end monoclonal antibody production process
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Date
2024
Authors
Pradeep, Rithwik
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Publisher
University College Cork
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Abstract
Monoclonal antibodies (mAbs) represent the dominant modality of biological therapeutics currently marketed. The manufacture of monoclonal antibodies consists of a series of processing steps including upstream cell culture, downstream purification and formulation and a final step of drug product manufacture. In this thesis, a small-scale end-to-end production process was developed for two monoclonal antibodies namely an anti-TNF Alpha mAb and cNISTmAb whose cell lines were sourced from commercial and public bodies respectively. A cell culture process for both cell lines was successfully developed including the creation of working cell banks (WCB). Titers were optimised, for both cell lines through a series of media, bioreactor and cell density modifications. In addition, the critical quality attributes of potency and purity were assessed via cell-based assays and SDS-PAGE respectively, that were designed and developed as part of this thesis. The potency of anti-TNF Alpha mAbs produced was assessed through two distinct in-vitro cell-based assays developed using HEK and L929 reporter cell lines and determined to be functional and more potent than a commercially available anti-TNF Alpha antibody The stability of cNISTmAb was also analysed across a range of temperature conditions from -20°C to 40°C after 50 days and also post-lyophilisation, with stability maintained at room temperature. Additional modifications of cNISTmAb were performed in an attempt to yield antibody fragments through pepsin digestion with partial success through pepsin digestion to yield F(ab’)2 fragments. This work demonstrates the successful production of two monoclonal antibodies and fragments but also sets the stage for further process improvement and enhanced characterisation to fully define and analyse an endto-end monoclonal antibody production system
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Keywords
Cell culture , Monoclonal antibody , Purification
Citation
Pradeep, R. 2024. Development of an end-to-end monoclonal antibody production process. MRes Thesis, University College Cork.