Employing QbD strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodies
Abstract Quality by Design (QbD) is one of the most important tools for the implementation of Process Analytical Technology (PAT) in biopharmaceutical production. For optimal characterization of a monoclonal antibody (mAb) upstream process a stepwise approach was implemented. The upstream was divide...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley-VCH
2023-02-01
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| Series: | Engineering in Life Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/elsc.202200056 |
| _version_ | 1797934821956124672 |
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| author | Ole Jacob Wohlenberg Carlotta Kortmann Katharina V. Meyer Thomas Scheper Dörte Solle |
| author_facet | Ole Jacob Wohlenberg Carlotta Kortmann Katharina V. Meyer Thomas Scheper Dörte Solle |
| author_sort | Ole Jacob Wohlenberg |
| collection | DOAJ |
| description | Abstract Quality by Design (QbD) is one of the most important tools for the implementation of Process Analytical Technology (PAT) in biopharmaceutical production. For optimal characterization of a monoclonal antibody (mAb) upstream process a stepwise approach was implemented. The upstream was divided into three process stages, namely inoculum expansion, production, and primary recovery, which were investigated individually. This approach enables analysis of process parameters and associated intermediate quality attributes as well as systematic knowledge transfer to subsequent process steps. Following previous research, this study focuses on the primary recovery of the mAb and thereby marks the final step toward a holistic characterization of the upstream process. Based on gained knowledge during the production process evaluation, the cell viability and density were determined as critical parameters for the primary recovery. Directed cell viability adjustment was achieved using cytotoxic camptothecin in a novel protocol. Additionally, the cell separation method was added to the Design of Experiments (DoE) as a qualitative factor and varied between filtration and centrifugation. To assess the quality attributes after cell separation, the bioactivity of the mAb was analyzed using a cell‐based assay and the purity of the supernatant was evaluated by measurement of process related impurities (host cell protein proportion, residual DNA). Multivariate data analysis of the compiled data confirmed the hypothesis that the upstream process has no significant influence on the bioactivity of the mAb. Therefore, process control must be tuned towards high mAb titers and purity after the primary recovery, enabling optimal downstream processing of the product. To minimize amounts of host cell proteins and residual DNA the cell viability should be maintained above 85% and the cell density should be controlled around 15 × 106 cells/ml during the cell removal. Thereby, this study shows the importance of QbD for the characterization of the primary recovery of mAbs and highlights the useful implementation of the stepwise approach over subsequent process stages. |
| first_indexed | 2024-04-10T18:04:56Z |
| format | Article |
| id | doaj.art-67c53fa960d54786ac1f684a20ecd154 |
| institution | Directory Open Access Journal |
| issn | 1618-0240 1618-2863 |
| language | English |
| last_indexed | 2024-04-10T18:04:56Z |
| publishDate | 2023-02-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Engineering in Life Sciences |
| spelling | doaj.art-67c53fa960d54786ac1f684a20ecd1542023-02-02T13:23:41ZengWiley-VCHEngineering in Life Sciences1618-02401618-28632023-02-01232n/an/a10.1002/elsc.202200056Employing QbD strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodiesOle Jacob Wohlenberg0Carlotta Kortmann1Katharina V. Meyer2Thomas Scheper3Dörte Solle4Leibniz Universität Hannover Institut für Technische Chemie Hannover GermanyLeibniz Universität Hannover Institut für Technische Chemie Hannover GermanyLeibniz Universität Hannover Institut für Technische Chemie Hannover GermanyLeibniz Universität Hannover Institut für Technische Chemie Hannover GermanyLeibniz Universität Hannover Institut für Technische Chemie Hannover GermanyAbstract Quality by Design (QbD) is one of the most important tools for the implementation of Process Analytical Technology (PAT) in biopharmaceutical production. For optimal characterization of a monoclonal antibody (mAb) upstream process a stepwise approach was implemented. The upstream was divided into three process stages, namely inoculum expansion, production, and primary recovery, which were investigated individually. This approach enables analysis of process parameters and associated intermediate quality attributes as well as systematic knowledge transfer to subsequent process steps. Following previous research, this study focuses on the primary recovery of the mAb and thereby marks the final step toward a holistic characterization of the upstream process. Based on gained knowledge during the production process evaluation, the cell viability and density were determined as critical parameters for the primary recovery. Directed cell viability adjustment was achieved using cytotoxic camptothecin in a novel protocol. Additionally, the cell separation method was added to the Design of Experiments (DoE) as a qualitative factor and varied between filtration and centrifugation. To assess the quality attributes after cell separation, the bioactivity of the mAb was analyzed using a cell‐based assay and the purity of the supernatant was evaluated by measurement of process related impurities (host cell protein proportion, residual DNA). Multivariate data analysis of the compiled data confirmed the hypothesis that the upstream process has no significant influence on the bioactivity of the mAb. Therefore, process control must be tuned towards high mAb titers and purity after the primary recovery, enabling optimal downstream processing of the product. To minimize amounts of host cell proteins and residual DNA the cell viability should be maintained above 85% and the cell density should be controlled around 15 × 106 cells/ml during the cell removal. Thereby, this study shows the importance of QbD for the characterization of the primary recovery of mAbs and highlights the useful implementation of the stepwise approach over subsequent process stages.https://doi.org/10.1002/elsc.202200056CHODoEmAbPATQbD |
| spellingShingle | Ole Jacob Wohlenberg Carlotta Kortmann Katharina V. Meyer Thomas Scheper Dörte Solle Employing QbD strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodies Engineering in Life Sciences CHO DoE mAb PAT QbD |
| title | Employing QbD strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodies |
| title_full | Employing QbD strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodies |
| title_fullStr | Employing QbD strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodies |
| title_full_unstemmed | Employing QbD strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodies |
| title_short | Employing QbD strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodies |
| title_sort | employing qbd strategies to assess the impact of cell viability and density on the primary recovery of monoclonal antibodies |
| topic | CHO DoE mAb PAT QbD |
| url | https://doi.org/10.1002/elsc.202200056 |
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