Development of serum-free medium supplemented with hydrolysates for the production of therapeutic antibodies in CHO cell cultures using design of experiments.
ABSTRACT An efficient method of formulating serum-free medium (SFM) for production of therapeutic antibodies by recombinant CHO (rCHO) cells was developed using two rCHO cell lines producing a therapeutic antibody. In this method, ten kinds of SFM were prepared by supplementing the basal SFM with statistically designed mixtures (total 5 g L(-1)) of three non-animal-derived hydrolysates: yeastolate, soy hydrolysate, and wheat gluten hydrolysate. When the two rCHO cell lines were cultivated, the mixtures of soy hydrolysate and wheat gluten hydrolysate showed a positive effect on cell growth. On the other hand, the mixtures including a high portion of yeastolate significantly enhanced specific antibody productivity. To reconstitute the mixture ratios of the three hydrolysates for high growth and antibody production, the effect of each medium was analyzed by the statistical program Design-Expert(R). The resulting medium gave a 1.9-3.3-fold increase in the maximum antibody concentration, compared to the basal SFM. Taken together, the supplementation of hydrolysates to the basal SFM with the help of statistical analysis is an efficient means of developing SFM for therapeutic antibody production by rCHO cells.
Article: Chromatographic fractionation of yeast extract: A strategy to identify physicochemical properties of compounds promoting CHO cell culture[show abstract] [hide abstract]
ABSTRACT: The study proposes to get a better knowledge of the physicochemical properties of yeast extract (YE) molecules involved in the improvement of CHO cell growth and to reduce YE complexity without losing positive effects. Various chromatographic processes were implemented for fractionation of a nanofil-trated YE (nYE). The nYE was first fractionated by one-step preparative chromatography, either with anion exchange (AEC), hydrophobic interaction (HIC) or size exclusion (SEC) methods. After analysis of its main components, each fraction was added in a control chemically defined medium to assess its impact on CHO cell growth. Results mainly underlined that AEC was the most selective separation process to purify nYE in one step without decreasing cell growth promoting effect. A three-step chromatographic process including successive AEC, HIC, and SEC was then developed to refine the physicochemical prop-erties of nYE compounds. Among fractions that triggered similar cell growth promoting effect than nYE, one also improved IgG specific production. It mainly included cationic and hydrophilic peptides with a great proportion of lysine and arginine, low quantities of polysaccharides and no nucleic acids. Thus, this study allowed us to deepen the YE contribution to animal cell culture as well as to evaluate fractionation strategies to simplify such a complex mixture.Process Biochemistry 01/2012; 47:1178-1185. · 2.44 Impact Factor