Production of recombinant therapeutic proteins because of their capacity for correct protein folding, assembly and post-translational modification. Inside the search of an effective method for the production of a recombinant protein employing animal cell culture, we investigated the effects of unique treatment including fetal calf serum concentration, glycerol and culture temperature on a Chinese hamster ovary (CHO) cell line on the production of recombinant human development hormone (rhGH) and recombinant Chinese hamster ovary (rCHO) viability. The GH production was assessed making use of ELISA and western blotting methods and cell viability was determined by flow cytometry. The production of recombinant protein elevated by 2-fold when stimulatory chemical for instance glycerol was added in two stages, 1st cells have been cultured with no glycerol for any time period to be able to obtain adequate cell density and then glycerol was added to achieve higher distinct productivity.. In addition, glycerol addition improved cell viability. Low culture temperature (under 37 ) led to enhanced cellular productivity in the rhGH by 3-fold but decreased cell viability. These findings indicate that quite straightforward variables like culture temperature and addition of simple chemical compounds may perhaps result in the improvement of industrial procedure for the production of recombinant proteins for example rhGH.Keywords: Chinese hamster ovary; rhGH productivity; Culture temperature; Glycerol; FCS; Cell viability INTRODUCTION Recombinant therapeutic proteins have changed the face of contemporary medicine and they continue to supply new and powerful therapies for various illnesses ranging from cancers to infertility (1). About 60-70 of all recombinant pharmaceutical proteins are made in mammalian cells. Quite a few of these proteins are expressed in immortalized Chinese hamster ovary (CHO) cells, but other cell lines, such as mouse myeloma (NSO), infant hamster kidney (BHK) and human embryonic kidney (HEK293) cells have gained approval for recombinant protein production (2). CHO cell culture is becoming increasingly significant for production of recombinant therapeutic proteins with respect to fast growth price, high production of recombinant proteins, and high stability of foreign gene expression (3). Growing recombinant protein yields, and thereby, reducing production charges are a major*Corresponding author: S.Buspirone H. Zarkesh-Esfahani Tel. 0098 311 7932473, Fax. 0098 311 7932456 E mail: s.Toceranib phosphate h.PMID:23983589 [email protected] targets (4). Since the productivity of mammalian cells is low as compared with that of prokaryotic hosts, you will find some tactics so as to increase recombinant protein production in CHO cells, like regulation of culture temperature (5), addition of various chemical substances (six), bioreactor engineering (7), and inhibition of cell apoptosis (8, 9). Culture temperature is amongst the most significant parameters that might be optimized for growing recombinant protein yields. Animal cells for example CHO are the most typically cultured at 37 to imitate the body environment. Since the culture temperature, at the same time as the medium pH (10-12) and dissolved oxygen (DO) (12-14) would impact some vital cellular situations for instance growth, viability and protein synthesis, they should be studied to figure out the appropriate method for protein production (15). Glycerol is among the chemical elements in medium that aids in forming a solvent shell about a proteinM. Rezaei et al. / RPS 2013; 8(three): 211-molecule as.
