Evaluation of the New Protein-Free Powder Medium (MDSS1-Powder) for the Production of Monoclonal Antibodies

Abstract

The use of serum-free media (SFM) is an important improvement in animal cell biotechnology. Classically, these media are commercialized in a liquid form which poses several technical problems for their distribution to industrial users. Therefore, the SFM MDSS1 which is originally a liquid medium, was further developed in order to obtain a full powder medium.

The evaluation of this medium consisted in comparing cell growth of three mouse-mouse hybridomas (I.13.17, 9E10, TB/C3 derived from 653, SP2/0, and NS1 myeloma cells, respectively) and monoclonal antibody (MAB) production in the original liquid MDSS1 as well as in the powder MDSS1 by using static batch and continuous stirred tank reactor cultures. In continuous cultures, the specific growth rates were equally or better in the powder MDSS1 (0.017-0.039/h) than in the originally liquid MDSS1 (0.0160.026/h). The steady state cell densities varied for the cell lines tested between 2 and 3*10“6 c/ml mainly as a result of differences in the cell retention by the spinfilters used. The specific IgG production rates varied very much in dependence of the cell line used. For 9E10 we could state a considerable increase from 1.2 to 6.6 pg IgG/c.h, for I.13.17 no statistically significant difference was stated, and for TB/C3 a three-fold reduction from 0.78 to 0.26 pg IgG/c.h was stated when the powder MDSS1 was used instead of the liquid formulation of MDSS1. In absolute values, IgG concentrations ranged from 50 to 300 mg/l. These results signify that both types of MDSS1 are equally useful for the production of MAB in reactor cultures. However, it is also evident that different hybridoma cell lines perform differently in both SFM and that the choice of the SFM depends finally on the cell line used.

one serum-free powder medium was commercialized up to know. Effectively Marcor’s LC115 serum-free medium is sold as a powder, however, we had a lot of difficulties to grow hybridoma cells on this medium even after extensive trials of adaptation. Only for one mouse hybridoma cell line we were able to conduct CSTR cultures (not shown).

In conclusion, this evaluation showed that protein-free powder media can successfully be used for the proliferation of mouse hybridoma cells and the production of MAB using various culture systems.