About this Biochemistry, Genetics and Molecular Biology article

Temperature control of growth and productivity in mutant Chinese hamster ovary cells synthesizing a recombinant protein by Nigel Jenkins; Alison Hovey is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

The use of a temperature switch to control the growth and productivity of temperature-sensitive (ts) mutants was investigated to extend the productive life span of recombinant Chinese hamster ovary (CHO) cells in batch culture. Bromodeoxyuridine was used at 39°C to select mutagenized CHO-K1 cells, which resulted in the isolation of 31 temperature-sensitive mutants that were growth inhibited at 39°C. Two of these mutants were successfully transfected with the gene for tissue inhibitor of metalloproteinases (TIMP) using glutamine synthetase amplification, and a permanent recombinant cell line established (5GI-Bl) that maintains the ts phenotype. Continuous exposure to the nonpermissive temperature (npt) of 39°C led to a rapid decline in cell viability. However, a temperature regime using alternating incubations at 34°C and 39°C arrested the 5G1-B1 cells while retaining a high cell viability for up to 170 h in culture. The specific production rate of the growtharrested cells was 3-4 times that of control cultures maintained at a constant 34°C over the crucial 72-130-h period of culture, which resulted in a 35% increase in the maximum product yield. Glucose uptake and lactate productio

Author

Nigel Jenkins; Alison Hovey

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); Wiley - V C H Verlag GmbbH & Co.; Wiley (ISSN 0006-3592)

Published

1993

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)