Quantitative analysis of transcription and translation in gene amplified Chinese hamster ovary cells on the basis of a kinetic model.

Cytotechnology
Martin SchröderPeter Friedl

Abstract

The elevation of expression levels for secreted glycoproteins by gene amplification in mammalian cells shows a saturation behavior at high levels of gene amplification. At high expression levels a drop in the secretion efficiency for the recombinant protein occurs (Schröder and Friedl, 1997), coinciding with the appearance of misfolded protein in the cell. In this communication we investigated whether additional limitations exist at the levels of transcription and translation. Four Chinese hamster ovary (CHO) cell lines expressing different amounts of human antithrombin III (ATIII) were used as a model system. A tenfold increase in the ATIII cDNA copy number from the lowest to the highest producing cell line coincided with a 38-fold increase in ATIII mRNA levels, and an 80-fold increase in the amount of intracellular ATIII levels. The data was analyzed using a simple kinetic model. The following conclusions were derived: I. The transcriptional activity for the recombinant protein is not saturated. II. Translation itself is not saturated either, but may be downregulated as secretion efficiency drops. III. Two explanations for the previously reported drop in secretion efficiency for the recombinant protein with increasing express...Continue Reading

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Citations

Sep 22, 2007·Biotechnology Letters·Martin Schröder
Nov 13, 2008·Cytotechnology·Saeed U Khan, Martin Schröder
Mar 1, 2002·Biotechnology and Bioengineering·Martin SchröderPeter Friedl
Feb 22, 2014·Applied Microbiology and Biotechnology·David ReinhartRenate Kunert
Jun 3, 2021·Cells·Anja J EngelGerhard Thiel

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