Short- and long-term effects on mAb-producing CHO cell lines after cryopreservation
| Autor: | Pynn Abigail Friederike Joyce, Inn H. Yuk, Rigzen P. S. Aulakh, Jayashree Subramanian, Mark Sanford, Parbir Grewal |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine medicine.drug_class Cell Clone (cell biology) CHO Cells Monoclonal antibody 01 natural sciences Cryopreservation 03 medical and health sciences Cricetulus Glutamate-Ammonia Ligase Methionine Sulfoximine 010608 biotechnology medicine Animals biology Chinese hamster ovary cell Antibodies Monoclonal Flow Cytometry Cell biology 030104 developmental biology medicine.anatomical_structure Cell culture biology.protein Antibody Intracellular Biotechnology |
| Zdroj: | Biotechnology Progress. 34:463-477 |
| ISSN: | 8756-7938 |
| DOI: | 10.1002/btpr.2599 |
| Popis: | Cryopreservation provides the foundation for research, development, and manufacturing operations in the CHO-based biopharmaceutical industry. Despite its criticality, studies are lacking that explicitly demonstrate that the routine cell banking process and the potential stress and damage during cryopreservation and recovery from thaw have no lasting detrimental effects on CHO cells. Statistics are also scarce on the decline of cell-specific productivity (Qp ) over time for recombinant CHO cells developed using the glutamine synthetase (GS)-based methionine sulfoximine (MSX) selection system. To address these gaps, we evaluated the impact of freeze-thaw on 24 recombinant CHO cell lines (generated by the GS/MSX selection system) using a series of production culture assays. Across the panel of cell lines expressing one of three monoclonal antibodies (mAbs), freeze-thaw did not result in any significant impact beyond the initial post-thaw passages. Production cultures sourced from cryopreserved cells and their non-cryopreserved counterparts yielded similar performance (growth, viability, and productivity), product quality (size, charge, and glycosylation distributions), and flow cytometric profiles (intracellular mAb expression). However, many production cultures yielded lower Qp at increased cell age: 17 of the 24 cell lines displayed ≥20% Qp decline after ∼2-3 months of passaging, irrespective of whether the cells were previously cryopreserved. The frequency of Qp decline underscores the continued need for understanding the underlying mechanisms and for careful clone selection. Because our experiments were designed to decouple the effects of cryopreservation from those of cell age, we could conclusively rule out freeze-thaw as a cause for Qp decline. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:463-477, 2018. |
| Databáze: | OpenAIRE |
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