Metabolically optimised BHK cell fed-batch cultures

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• 作者：Cruz ; Helder J. ; Moreira ; José ; L. ; Carrondo ; Manuel J.T.
• 关键词：Metabolic optimisation ; Metabolic shifts ; Fed-batch cultures ; BHK cells ; Fusion protein ; Tumour targeted therapy ; Glucose and glutamine consumption
• 刊名：Journal of Biotechnology
• 出版年：2000
• 出版时间：June 23, 2000
• 年：2000
• 卷：80
• 期：2
• 页码：109-118
• 全文大小：138 K

文摘

The aim of this work was the optimisation of a fed-batch culture by metabolic confinement of BHK21 cells producing an antibody/cytokine fusion protein with potential application in tumour-targeted therapy. Previous results showed that at very low nutrient concentrations, a metabolic shift towards more efficient metabolic pathways occurs. The application of those results in the optimisation of a fed-batch culture resulted in higher cell growth (0.020 vs. 0.016 h⁻¹) and cell viability, higher maximum cell concentration (2.5 vs. 1.1×10⁶ cell ml⁻¹), longer culture span (17 versus nine days) and higher product titre (60 % increase), in relation to batch culture. This was achieved by maintaining glucose at 0.3 mM and glutamine at 0.2 mM through the addition of a concentrated solution based on the estimations of future nutrient consumption and growth rates through off line measurements. The production of toxic metabolites such as lactate and ammonia was reduced, especially the lactate production, which was markedly decreased due to the metabolic confinement of the cells. In conclusion, it was possible to increase the final titre of the recombinant antibody/cytokine fusion protein by confining the metabolism of the cells to an energetically more efficient state.