91桃色

Abstract

Today, almost 60-70% of recombinant proteins are successfully produced in CHO cells. The optimization of cell culture conditions for growth and productivity of recombinant Chinese hamster ovary (CHO) cells is a critical step in biopharmaceutical manufacturing. Transient expression of the protein (Transient Gene Expression) TGE is the preferred way to express recombinant proteins in a short timeframe and at a scale suitable for pre-clinical testing. In the present study, the effect of optimization of transfection condition in a serum free medium CD DG44 in a Shaking incubator with CHO DG44 cells in suspended condition was evaluated . A range of values for DNA, PEI and Cell concentration were studied to find the optimal values regarding transfection efficiency. The results showed that the optimized values were 1.5 μg/106 cells of trasfection reagent (PEI), 0.5*106 cells for starting cell densities and 2 μg/106 cells of DNA with 28.71, 38.68 and 52.8% transfection efficiency respectively. Furthermore, based on these TGE optimized condition effect of feeding with 6 different Soy and Caseine peptones on transfection efficiency was evaluated The results of peptone feeding with peptones Caseine Tryptone N1, Soy Peptone A2Sc and Soy peptone E110 showed 59.88.%, 58.28% and 68.66% transfection efficiency with 15.44, 12.35 and 28.55% increase compared to previous optimized process. These data shows that the appropriate feeding strategy is capable of increasing transfection efficiency and is a promising approach for optimizing TGE process.

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