An experimental factorial design was developed to study influence of seed cell density, agitation rate and culture condition(open or closed system) in order to maximize the expression of recombinant protein transfected in CHO cell line. A 2 3 experimental design included those independent variables was running and total protein expression and specific protein expression were measured as dependent variables. A low and high level was established for each variable assuming working experience and literature review. There were running 8 experiments keeping similar culture conditions except variables under study. Multiple lineal regression model was fitted to the data and statistic parameters suggest a good correlation between independent and dependent variables Results show that three independent variables have a statistic significance in the expression of target protein but not interaction between them. These results are very useful in order to establish culture condition at large scale production of biopharmaceuticals and for validation purposes.
INTRODUCTIONRoller bottle systems have been used for over 50 years in pharmaceutical and biochemical process. (Liu YL et al, 2003, Tsao EI et al, 1992 Widespread use of the roller bottle is due to several reasons like simplicity and scalability of the technology, reduced developmental timelines, constant fluid-gas contact and capability to maintain sterile conditions. (Muzzio et al, 1999) An experimental factorial design was developed to study influence of seed cell density, agitation rate and culture condition(open or closed system) in order to maximize the expression of recombinant protein transfected in CHO cell line.
MATERIALS AND MÉTHODS.The CHO cell line used is derived from wild type (ATCC CRL-9096) and transformed to express heterologous protein in our Institute (Center for Genetic Engineering and Biotechnology). The cells were cultivated in Dulbecco´s Modified Eagle Medium (D-MEM) containing 5 % of fetal bovine serum (FBS) supplemented with 10 µg/mL gentamicine. Cells were kept at 37°C. Medium exchange was done u u Ha
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