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Producer vs. parental cell - metabolic changes and burden upon α1-antitrypsin production in AGE1.HN(®).

Niklas J, Priesnitz C, Sandig V, Rose T, Heinzle E - BMC Proc (2011)

View Article: PubMed Central - HTML - PubMed

Affiliation: Biochemical Engineering Institute, Saarland University, 66123 Saarbrücken, Germany.

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The questions addressed were (i), which changes occur in cell growth and metabolism upon A1AT production and (ii), what are specific cellular properties distinguishing the producer clone AGE1.HN.AAT from the parental cell population... Glycine and glutamate production was higher in AGE1.HN.AAT whereas uptake of arginine and aspartate as well as alanine production were lower... Glutamine was consumed in the cultivations of both cell lines at ~4 days which resulted also in reduced growth... The differences that can be seen in the metabolism upon production of the glycoprotein were explained by using an appropriate model of the anabolic demand needed to produce active A1AT... The whole cellular production process of a glycoprotein includes its expression (transcription), synthesis of the amino acid chain (translation), posttranslational modification in endoplasmic reticulum (ER) and Golgi apparatus (Golgi) and secretion of the mature protein (Figure 1)... These results indicate that one could expect an increase in the production of glutamate and glycine with increasing intracellular nucleotide demand which was observed for AGE1.HN.AAT; this is caused by differences in cellular composition of the producer, partly originating from recombinant A1AT production... The differences that were found in the biomass composition between both cell lines were included in a metabolic network model and intracellular metabolic fluxes were calculated for both cell lines... Fluxes in central energy metabolism (glyolysis, TCA cycle) were similar... Differences were observed in C1 and glutamate metabolism including changes in activities of several transaminases... The observed changes in metabolic flux reflect the anabolic differences between producer and parental cells... This improved understanding of the metabolism and cellular changes during glycoprotein production supports the identification of targets for further improvement of (i) cell line, e.g., by genetic modifications and (ii) cultivation process, e.g., by improved feeding strategies... The presented data indicate that nucleotide and lipid metabolism might be interesting targets for further engineering of the AGE1.HN cell line.

No MeSH data available.


Schematic presentation of the glycoprotein production process in mammalian cells.
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Figure 1: Schematic presentation of the glycoprotein production process in mammalian cells.

Mentions: The differences that can be seen in the metabolism upon production of the glycoprotein were explained by using an appropriate model of the anabolic demand needed to produce active A1AT. The whole cellular production process of a glycoprotein includes its expression (transcription), synthesis of the amino acid chain (translation), posttranslational modification in endoplasmic reticulum (ER) and Golgi apparatus (Golgi) and secretion of the mature protein (Figure 1). Metabolite demand for A1AT production was finally simulated. These results indicate that one could expect an increase in the production of glutamate and glycine with increasing intracellular nucleotide demand which was observed for AGE1.HN.AAT; this is caused by differences in cellular composition of the producer, partly originating from recombinant A1AT production.


Producer vs. parental cell - metabolic changes and burden upon α1-antitrypsin production in AGE1.HN(®).

Niklas J, Priesnitz C, Sandig V, Rose T, Heinzle E - BMC Proc (2011)

Schematic presentation of the glycoprotein production process in mammalian cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3285021&req=5

Figure 1: Schematic presentation of the glycoprotein production process in mammalian cells.
Mentions: The differences that can be seen in the metabolism upon production of the glycoprotein were explained by using an appropriate model of the anabolic demand needed to produce active A1AT. The whole cellular production process of a glycoprotein includes its expression (transcription), synthesis of the amino acid chain (translation), posttranslational modification in endoplasmic reticulum (ER) and Golgi apparatus (Golgi) and secretion of the mature protein (Figure 1). Metabolite demand for A1AT production was finally simulated. These results indicate that one could expect an increase in the production of glutamate and glycine with increasing intracellular nucleotide demand which was observed for AGE1.HN.AAT; this is caused by differences in cellular composition of the producer, partly originating from recombinant A1AT production.

View Article: PubMed Central - HTML - PubMed

Affiliation: Biochemical Engineering Institute, Saarland University, 66123 Saarbrücken, Germany.

AUTOMATICALLY GENERATED EXCERPT
Please rate it.

The questions addressed were (i), which changes occur in cell growth and metabolism upon A1AT production and (ii), what are specific cellular properties distinguishing the producer clone AGE1.HN.AAT from the parental cell population... Glycine and glutamate production was higher in AGE1.HN.AAT whereas uptake of arginine and aspartate as well as alanine production were lower... Glutamine was consumed in the cultivations of both cell lines at ~4 days which resulted also in reduced growth... The differences that can be seen in the metabolism upon production of the glycoprotein were explained by using an appropriate model of the anabolic demand needed to produce active A1AT... The whole cellular production process of a glycoprotein includes its expression (transcription), synthesis of the amino acid chain (translation), posttranslational modification in endoplasmic reticulum (ER) and Golgi apparatus (Golgi) and secretion of the mature protein (Figure 1)... These results indicate that one could expect an increase in the production of glutamate and glycine with increasing intracellular nucleotide demand which was observed for AGE1.HN.AAT; this is caused by differences in cellular composition of the producer, partly originating from recombinant A1AT production... The differences that were found in the biomass composition between both cell lines were included in a metabolic network model and intracellular metabolic fluxes were calculated for both cell lines... Fluxes in central energy metabolism (glyolysis, TCA cycle) were similar... Differences were observed in C1 and glutamate metabolism including changes in activities of several transaminases... The observed changes in metabolic flux reflect the anabolic differences between producer and parental cells... This improved understanding of the metabolism and cellular changes during glycoprotein production supports the identification of targets for further improvement of (i) cell line, e.g., by genetic modifications and (ii) cultivation process, e.g., by improved feeding strategies... The presented data indicate that nucleotide and lipid metabolism might be interesting targets for further engineering of the AGE1.HN cell line.

No MeSH data available.