Hybridomas production levels

Optimal fermentation parameters have been well established and air-lift, stirred tank, and hollow fibre systems have all been used. At commercial scale, fermentation volumes in excess of 1000 litres can be used, which can yield 100 g or more of final product. While hybridoma growth is straightforward, production levels of antibody can be quite low compared with ascites-based production systems. Typically, fermentation yields antibody concentrations of 0.1-0.5 mg/ml. Removal of cells from the antibody-containing media is achieved by centrifugation or filtration. An ultrafiltration step is then normally undertaken in order to concentrate the filtrate by up to 20-fold. 

The most important technique for perfusion culture methods is to separate the concentrated cells and conditioned medium from the suspended culture broth. As noted above, the separation methods chiefly used are filtration with tubular and flat membranes as well as ceramic macroporous filters. These membrane reactors can be employed for both anchorage-dependent and suspension growing cells. Static maintenance type systems are commercially available for disposable reactors, and small size unit reactors from 80 ml to 1 liter are used for continuous production of monoclonal antibodies with hybridoma cells. The maintainable cell densities are about 10 -10 cells/ ml which is essentially mouse ascites level. However, in these systems, the cell numbers cannot be counted directly because the cells adhere to membranes or hollow fibers. Therefore, the measurement of cell density must use indirect methods. Such indirect methods include the assaying of the quantities of glucose consumption and the accumulation of lactate. The parameters of scale-up have not yet been established for these static methods. 

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