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Process analysis of reduced specific productivity of TNFR-Fc in Chinese hamster ovary cells at high cell density

Paper ID Volume ID Publish Year Pages File Format Full-Text
34912 45059 2011 8 PDF Available
Title
Process analysis of reduced specific productivity of TNFR-Fc in Chinese hamster ovary cells at high cell density
Abstract

Specific TNFR-Fc productivity (qTNFR-Fc) in CHO cells was found to decrease by 68% at a cell density of 6 × 106 cells/mL compared to that at 2 × 106 cells/mL. In order to unravel the reasons for the reduced qTNFR-Fc at high cell density, an in-depth study of physiological factors, gene transcription and post-translational process was carried out. Physiological factors including cell biomass, intracellular protein and cell cycle distribution were not affected by cell density. Transcriptional analysis revealed that TNFR-Fc mRNA level at 2 × 106 cells/mL was almost 2-fold higher than that at 6 × 106 cells/mL. Moreover, rather than mRNA stability, the slower transcriptional rate led to the decreased mRNA level. By analyzing the post-translational process, we found that the protein assembly rate and transport rate from the endoplasmic reticulum to the Golgi at high cell density were significantly lower than those at low cell density. Therefore, the decreased qTNFR-Fc at high cell density was correlated with the reduction in both mRNA level and post-translational processing rate. It was further found that the decreased mRNA level and post-translational processing rate might be linked to the apparent increase in CO2 partial pressure (pCO2) at high cell density, which had resulted in the reduction in intracellular pH in CHO cells.

Keywords
Cell density; CHO cells; TNFR-Fc; Specific productivity; Process analysis
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Process analysis of reduced specific productivity of TNFR-Fc in Chinese hamster ovary cells at high cell density
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 46, Issue 7, July 2011, Pages 1492–1499
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us