Systematic improvement of a chemically-defined protein-free medium for hybridoma growth and monoclonal antibody production

Effects of Supplementation of Various Medium Components on Chinese Hamster Ovary Cell Cultures Producing Recombinant Antibody

“…Since it has been reported that vitamin depletion can limit cell growth and supplementation of some vitamins enhance either cell growth or recombinant antibody production (Stoll et al 1996;Ducommun et al 2001;Ishaque and AlRubeai 2002), it was decided to double the concentration of each vitamin in the basal protein-free medium to prevent any vitamin depletion. Thirteen water-soluble vitamins (D-biotin, D-calcium pantothenate, choline chloride, cyanocobalamine, folic acid, i-inositol, niacinamide, pyridoxalÁHCl, pyridoxineÁHCl, riboflavin, thiamineÁHCl, ascorbic acid, folinic acid) present in the basal protein-free medium were considered, and their concentration was changed individually.…”

“…In hybridoma cultivation, several kinds of amino acid such as glutamine, asparagines, and glycine are known to prevent cellular apoptosis (Stoll et al 1996;Ducommun et al 2001). Appropriately balanced supplementation of amino acid was shown to be very efficient to increase viable cells and final concentration of recombinant antibody (Stoll et al 1996;Ducommun et al 2001).…”

“…Also, various hormones, inorganic salts, and other chemical reagents are known to improve the cell growth and productivity of therapeutic proteins when they are added to the media (Morris and Schmid 2000;Sunstrom et al 2000). Also, several strategies for the fortification of basal media have been developed to improve culture longevity (Stoll et al 1996;Ducommun et al 2001).…”

“…Animal cells require various nutrients such as amino acids, vitamins, hormones, inorganic salts, and diverse undefined components to survive in vitro and produce the target therapeutic proteins efficiently. Because most of mammalian cells are very sensitive to their environmental change, balanced addition of limiting components with a detailed medium analysis take a critical portion in the modification of medium formulation (Stoll et al 1996;Ducommun et al 2001). Additional supplementation of amino acids and vitamins to serumfree or protein-free media has resulted in remarkable increases in viable cell density and final recombinant antibody production (Stoll et al 1996;Ducommun et al 2001;Takagi et al 2001;Ishaque and Al-Rubeai 2002).…”

“…Because most of mammalian cells are very sensitive to their environmental change, balanced addition of limiting components with a detailed medium analysis take a critical portion in the modification of medium formulation (Stoll et al 1996;Ducommun et al 2001). Additional supplementation of amino acids and vitamins to serumfree or protein-free media has resulted in remarkable increases in viable cell density and final recombinant antibody production (Stoll et al 1996;Ducommun et al 2001;Takagi et al 2001;Ishaque and Al-Rubeai 2002). Also, various hormones, inorganic salts, and other chemical reagents are known to improve the cell growth and productivity of therapeutic proteins when they are added to the media (Morris and Schmid 2000;Sunstrom et al 2000).…”