Bioreactor scale up and protein product quality characterization of piggyBac transposon derived CHO pools
| Autor: | Dawn L. Norris, Yashas Rajendra, Sowmya Balasubramanian, Gavin C. Barnard, James R. Swartling, Neil A. McCracken, Robert B. Peery, Christopher C. Frye |
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| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine medicine.drug_class Pilot Projects CHO Cells Biology Monoclonal antibody Protein Engineering 01 natural sciences 03 medical and health sciences Bioreactors Cricetulus 010608 biotechnology Bioreactor medicine Animals Bioprocess Gene Cell Proliferation Messenger RNA Chinese hamster ovary cell Antibodies Monoclonal Molecular biology Recombinant Proteins Up-Regulation 030104 developmental biology Biopharmaceutical Biochemistry Cell culture Batch Cell Culture Techniques DNA Transposable Elements Biotechnology |
| Zdroj: | Biotechnology progress. 33(2) |
| ISSN: | 1520-6033 |
| Popis: | Chinese hamster ovary (CHO) cells remain the most popular host for the production of biopharmaceutical drugs, particularly monoclonal antibodies (mAbs), bispecific antibodies, and Fc-fusion proteins. Creating and characterizing the stable CHO clonally-derived cell lines (CDCLs) needed to manufacture these therapeutic proteins is a lengthy and laborious process. Therefore, CHO pools have increasingly been used to rapidly produce protein to support and enable preclinical drug development. We recently described the generation of CHO pools yielding mAb titers as high as 7.6 g/L in a 16 day bioprocess using piggyBac transposon-mediated gene integration. In this study, we wanted to understand why the piggyBac pool titers were significantly higher (2-10 fold) than the control CHO pools. Higher titers were the result of a combination of increased average gene copy number, significantly higher messenger RNA levels and the homogeneity (i.e. less diverse population distribution) of the piggyBac pools, relative to the control pools. In order to validate the use of piggyBac pools to support preclinical drug development, we then performed an in-depth product quality analysis of purified protein. The product quality of protein obtained from the piggyBac pools was very similar to the product quality profile of protein obtained from the control pools. Finally, we demonstrated the scalability of these pools from shake flasks to 36L bioreactors. Overall, these results suggest that gram quantities of therapeutic protein can be rapidly obtained from piggyBac CHO pools without significantly changing product quality attributes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:534-540, 2017. |
| Databáze: | OpenAIRE |
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