Modification of glucose and glutamine metabolism in hybridoma cells through metabolic engineering.
The present work describes the genetic modification of a hybridoma cell line with the aim to change its metabolic behaviour, particularly reducing the amounts of ammonia and lactate produced by the cells. The cellular excretion of ammonia was eliminated by transfection of a cloned glutamine synthetase gene. The metabolic characterisation of the transformed cell line includes the analysis of the changes introduced in its intracellular metabolic fluxes by means of a stoichiometric model. Furthermore, the reduction of lactate accumulation was attempted through an antisense mRNA approach, aiming to generate a rate limiting step in the glycolytic pathway, thus lowering the glucose consumption rate. The physiological results obtained with the transformed cells are discussed. A maximum reduction of about 47% in the glucose consumption rate was obtained for one of the transformations. However a main drawback was the lack of stability of the transformed cells.
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Departament d'Enginyeria Química, Facultat de Ciències, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain.
Prats
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Cairó
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Azorín
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Cornudella
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Gòdia
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