Influence of the number of animals on the production of monoclonal antibody CB.Hep-1 by the Ascites method

The ascites production method of inoculating hybridoma B cells in histocompatible mice can successfully produce monoclonal antibodies (MAb). The aim of this paper is to study the influence of the number of animals on MAb CB.Hep-1 production by the ascites production method. CB.Hep-1 mouse hybridoma cells were inoculated in different groups of animals (I, 750 mice; II, 1000 mice, III, 3500 mice; IV, 4000 mice and V, 6000 mice) previously irritated with mineral oil. The ascitic fluid was harvested through abdominal paracentesis and the results showed a marked influence of the number of animals on the amount of MAb CB.Hep-1 produced by the mice (I, 15.70 mg; II, 19.74 mg; III, 9.91 mg; IV, 8.46 mg; V, 5.30 mg). In conclusion, the concept of Reduction (3R concepts) was rigorously studied in this paper, determining the lowest number of inoculated animals needed for the highest amount of the MAb CB.Hep-1. Results indicate that the number of mice with tumors and the MAb CB.Hep-1 production decrease with the increase in the number of animals inoculated. Therefore between 50 000 and 250 000 animals could be spared each year if the number of animals was of ≤ 3500 per inoculation under the conditions studied here.