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Abstract Nowadays, Monoclonal antibodies (mAbs) have many potential applications in different fields such as diagnostic assays, therapeutics, downstream processing, and many other areas of biotechnological interest. mAbs accounted for 32 % of all revenues in the biotech market in 2008, and the worldwide market revenues for therapeutic and diagnostic antibodies are expected to reach about $ 26 billion in 2010. This has stimulated the need for the optimization of the existing production methods and the development of new ones. In the present study, several successful methods for optimization and scaling up production of mAb (IgG2a) from hybridoma cell line (OKT3) were performed: The first objective of this study was to adapt hybridoma cells to grow from serum enriched medium (DMEM supplemented with 20% FBS) to complete serum free medium (SMIF-6) using the technique of sequential adaptation at both T-flask and spinner flask levels, this is due to several problems which arise when FBS is used: its high cost, lack of homogeneity between different batches, and its high protein content, which make the downstream process more difficult. In this experiment kinetic studies have been carried out for hybridoma cell growth, metabolism, and mAb production during each adaptation step. Moreover, morphological observation of the cells cultured in different serum levels was carried out. It was noticed that compromise between cell growth and antibody production has been achieved; there are no apparent differences in cell growth in both media. Moreover, replacement of serum enriched media with serum free media enhanced mAb productivity. In addition, the gradual decrease in serum concentration in successive batches is not accompanied by observable change in the metabolic activity of the cells. Moreover, the cells cultured in different serum levels showed no cha |