| Autor: |
Shih-Hsie Pan, Samiksha Nayak, Zheng Jian Li, Michael C. Borys, Susan Egan, Yueming Qian, Ying Jing |
| Rok vydání: |
2012 |
| Předmět: |
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| Zdroj: |
Process Biochemistry. 47:69-75 |
| ISSN: |
1359-5113 |
| DOI: |
10.1016/j.procbio.2011.10.009 |
| Popis: |
The presence of aggregated forms of proteins can be problematic for therapeutics due to the potential for immunogenic and pharmacokinetic issues. Although downstream processing can remove the aggregated forms, inhibiting aggregate formation upstream during the cell culture stage could reduce the burden on downstream processing and potentially improve process yields. This study first examined the effects of environmental factors (temperature, pH, and dissolved oxygen) and medium components (bivalent copper ion, cysteine, and cystine) on the aggregation of two different recombinant fusion proteins expressed by Chinese hamster ovary (CHO) cells. Any strategy to reduce protein aggregation upstream during cell culture must also consider potential effects on critical upstream parameters such as cell growth, harvest titer, and protein sialylation levels. Manipulating the culture temperature shift and cystine concentration in the medium were both identified as effective and practical strategies for reducing protein aggregation in both CHO-cell expression systems. Furthermore, a combination of both strategies was more effective in reducing protein aggregation levels compared to either approach individually; and without any negative effects on harvest titer and protein sialylation. This study demonstrates a practical methodology for decreasing protein aggregation during upstream processing and emphasizes the importance of process understanding to ensure production of recombinant glycoprotein therapeutics with consistent product quality. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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