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Temperature control of growth and productivity in mutant Chinese-hamster ovary cells synthesizing a recombinant protein

Jenkins, Nigel, Hovey, Alison (1993) Temperature control of growth and productivity in mutant Chinese-hamster ovary cells synthesizing a recombinant protein. Biotechnology and Bioengineering, 42 (9). pp. 1029-1036. ISSN 0006-3592. (doi:10.1002/bit.260420903) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:22154)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided.
Official URL:
http://dx.doi.org/10.1002/bit.260420903

Abstract

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-degrees-C to select mutagenized CHO-K1 cells, which resulted in the isolation of 31 temperature-sensitive mutants that were growth inhibited at 39-degrees-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 (5G1-B1) that maintains the ts phenotype. Continuous exposure to the nonpermissive temperature (npt) of 39-degrees-C led to a rapid decline in cell viability. However, a temperature regime using alternating incubations at 34-degrees-C and 39-degrees-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 growth-arrested cells was 3-4 times that of control cultures maintained at a constant 34-degrees-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 production both decreased in arrested cells. Flow cytometric analysis indicated that 5G1-B1 cells arrested in the G1 or G0 phase of the cell cycle, and no major structural damage was caused to these cells by the alternating temperature regime. These results demonstrate that growth-arrested ts CHO cells have increased productivity compared to growing cultures and maintain viability for longer periods. The system offers the prospect of enhancing the productivity of recombinant mammalian cells grown in simple batch fermentors. (C) 1993 John Wiley & Sons, Inc.

Item Type: Article
DOI/Identification number: 10.1002/bit.260420903
Uncontrolled keywords: CHO - temperature-sensitive mutants - recombinant - growth arrest
Subjects: Q Science
Q Science > QR Microbiology
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: M. Nasiriavanaki
Date Deposited: 26 Jul 2009 19:06 UTC
Last Modified: 16 Nov 2021 10:00 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/22154 (The current URI for this page, for reference purposes)

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