Premium
Effect of shear stress on expression of a recombinant protein by Chinese hamster ovary cells
Author(s) -
Keane Julian T.,
Ryan David,
Gray Peter P.
Publication year - 2002
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10472
Subject(s) - chinese hamster ovary cell , shear stress , shear rate , cell culture , fetal bovine serum , critical resolved shear stress , chemistry , biology , microbiology and biotechnology , cell , materials science , biochemistry , composite material , rheology , genetics
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 μg/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. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 211–220, 2003.