Volume 7, Issue 3 (Aug 2019)                   Res Mol Med (RMM) 2019, 7(3): 0-0 | Back to browse issues page

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Robatjazi S M, Mahboudi S, Zeinoddini M. Production of Recombinant Denileukin Diftitox: Assessment in the Lab-scale Bioreactor. Res Mol Med (RMM). 2019; 7 (3)
Abstract:   (412 Views)
Introduction: Denileukin diftitox is a recombinant immunotoxin composed truncated diphtheria toxin fused to human interleukin 2 (DT389-IL2) and it has been considered as a drug candidate for protein therapy of lymphoma. The aim of this work is investigation of DT389-IL2 production using recombinant strain of Escherichia coli in the lab-scale bioreactor.
Material and methods: Firstly, the effect of chemical composition in culture medium (the complex and defined) was investigated in DT389-IL2 expression in E.coli BL21 (DE3) containing pET-IDZ plasmid in shack flask and lab-bioreactor culture. In order to enrichment carbon source, glucose 6-8 (g/l) was added to the complex culture medium. The composition of defined medium was enriched by adding the amino acids (valine, 0.0502; pheyalanine, 0.0132 lysine, 0.0184; aspartic acid, 0.016; serine, 0.0251 g/l). The growth cell was measured by optical density method and the expression yield was investigated by SDS-PAGE.
Results: The maximum growth rate was determined in modified complex culture medium containing glucose of 6 (g/l).  The bacteria growth in medium was achieved to the optical density of 5.5 in the shack flask culture, and under the same condition, the optical density of medium was increased to 13 in the bioreactor. Adding amino acids to the defined medium caused a significant enhancement on growth cell and expression of protein and the optical density of 2.95 and 6.1 were achieved in the shack-flask and bioreactor culture, respectability. The specific growth rate of bacteria in the complex medium and the defined medium was determined 0.89 and 0.47 h-1, respectability. 
Conclusion: The results shown, that adding the glucose carbon source to the complex medium and elemental enrichment of the defined medium by adding amino acids cased to increases biomass production. The growth cell rate in bioreactor by good controlling environmental conditions and proper mixing was higher compared with shack-flask culture.
Type of Study: Research | Subject: Biochemistry
Received: 2019/08/18

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