Effect of shear stress on expression of a recombinant protein by Chinese hamster ovary cells

Keane, Julian T., Ryan, David and Gray, Peter P. (2003) Effect of shear stress on expression of a recombinant protein by Chinese hamster ovary cells. Biotechnology and Bioengineering, 81 2: 211-220. doi:10.1002/bit.10472

Author Keane, Julian T.
Ryan, David
Gray, Peter P.
Title Effect of shear stress on expression of a recombinant protein by Chinese hamster ovary cells
Journal name Biotechnology and Bioengineering   Check publisher's open access policy
ISSN 0006-3592
Publication date 2003-01
Sub-type Article (original research)
DOI 10.1002/bit.10472
Volume 81
Issue 2
Start page 211
End page 220
Total pages 10
Editor Klee, Harry
Place of publication Hoboken
Publisher John Wiley & Sons Inc
Language eng
Subject 291804 Nanotechnology
290000 Engineering and Technology
Abstract A flow chamber was used to impart a steady laminar shear stress on a recombinant Chinese hamster ovary (CHO) cell line expressing human growth hormone (hGH). The cells were subjected to shear stress ranging from 0.005 to 0.80 N m(-2). The effect of shear stress on the cell specific glucose uptake, cell specific hGH, and lactate productivity rates were calculated. No morphological changes to the cells were observed over the range of shear stresses examined. When the cells were subjected to 0.10 N m(-2) shear in protein-free media without Pluronic F-68, recombinant protein production ceased with no change in cell morphology, whereas control cultures were expressing hGH at 0.35 mug/10(6) cells/h. Upon addition of the shear protectants, Pluronic F-68 (0.2% [w/v]) or fetal bovine serum (1.0% [v/v] FBS), the productivity of the cells was restored. The effect of increasing shear stress on the cells in protein-free medium containing Pluronic F-68 was also investigated. Cell specific metabolic rates were calculated for cells under shear stress and for no-shear control cultures performed in parallel, with shear stress rates expressed as a percentage of those obtained for control cultures. Upon increasing shear from 0.005 to 0.80 N m(-2), the cell specific hGH productivity decreased from 100% at 0.005 N m(-2) to 49% at 0.80 N m(-2) relative to the no-shear control. A concurrent increase in the glucose uptake rate from 115% at 0.01 N m(-2) to 142% at 0.80 N m(-2), and decreased lactate productivity from 92% to 50%, revealed a change in the yield of products from glucose compared with the static control. It was shown that shear stress, at sublytic levels in medium containing Pluronic F-68, could decrease hGH specific productivity. (C) 2002 Wiley Periodicals, Inc.
Keyword Biotechnology & Applied Microbiology
Chinese Hamster Ovary (cho)
Human Growth Hormone (hgh)
Recombinant Protein Productivity
Pluronic F-68
Shear Protectants
Vascular Endothelial-cells
Cultured Animal-cells
Bubble Interactions
Insect Cells
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Australian Institute for Bioengineering and Nanotechnology Publications
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Citation counts: TR Web of Science Citation Count  Cited 47 times in Thomson Reuters Web of Science Article | Citations
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Created: Mon, 13 Aug 2007, 13:20:11 EST