Hybridoma cell growth and anti-neuroblastoma monoclonal antibody production in spinner flasks using a protein-free medium with microcarriers

The main disadvantages of foetal calf serum as the world-wide common serum supplement for cell growth are its content of various proteins of variable concentrations between batches as well as its high cost. The use of serum-f ree and protein-free media is gradually becoming one of the goals of cell c ulture especially for standardizing culture conditions or for simple purifi cation of cell products like monoclonal antibodies. The mouse hybridoma cel ls 14/2/1 were cultivated either in protein-free UltraDOMA medium or in ser um-containing RPMI medium with and without microcarriers to generate high q uantities of monoclonal antibodies against neuroblastoma tumour cells. Cell growth rate, IgG production, viability, glucose and lactate concentrations , attachment rate and doubling time have been used as investigation criteri a. Modifications of culture procedures (static or stirred), inoculum densit y, and microcarrier concentration caused an improvement of monoclonal antib ody production. The kinetics of antibody synthesis was best in spinner cult ure with 2 ml of microcarriers in protein-free medium. These results of sho rt-term microcarrier culture in stirred spinner flasks indicate that IgG yi elds in protein-free medium 2.5-fold higher to those in serum-supplemented medium can be achieved. (C) 1999 Elsevier Science B.V. All rights reserved.