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J Immunol Methods. 1991 Dec 15;145(1-2):213-21.

Hybridoma antibody production in vitro in type II serum-free medium using Nutridoma-SP supplements. Comparisons with in vivo methods.

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Institut für Viruskrankheiten und Immunprophylaxe (IVI), Basel, Switzerland.


Six hybridoma clones (three producing anti-bovine viral diarrhoea antibodies and three producing anti-hog cholera virus antibodies) were studied with respect to their growth in type II serum-free medium. This was found to be successful in all cases when the serum-free medium was supplemented with 1% v/v Nutridoma, but only after adaptation over a small number of subcultures. Adapted hybridoma cultures could be grown in 250 ml spinner flasks, and the antibody-containing supernatants concentrated by tangential flow filtration. These were compared both with conventional cultures of the clones grown in serum-containing media and with ascites fluids generated using the latter type of culture. Adaptation to growth in the Nutridoma-based serum-free medium did not impair the capacity of the cells to produce and secrete antibodies, although their growth rate was slightly reduced when 250 ml spinner cultures were employed. The titre obtained from the ascites fluids were, as expected, higher than those from the tissue cultures, but contamination by other proteins, a significant problem in ascites fluids, did not present itself with the serum-free cultures. Consequently, 'serum-free' hybridoma antibodies could easily be concentrated in a relatively pure form, with respect to immunoglobulins present, having no need for affinity purification procedures. In addition, the acute in vivo problems of solid tumours and haemorrhage production could be avoided through the use of in vitro methods.

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