Simple Production and Purification of Monoclonal Antibodies in Serum-Free Medium

Authors: AYNUR BAŞALP, BEYAZIT ÇIRAKOĞLU, ENGIN BERMEK

Abstract: Although serum is a main source of essential nutrients and growth factors, its use creates highly complex and poorly defined media. The use of serum-free media is more efficient than the use of media containing serum especially when the purification and characterization of antibodies are considered. In this study, we developed a serum-free medium system for cultivation of hybrid cells producing monoclonal antibodies specific for Tobacco Mosaic Virus (1F5) and 17\beta-estradiol (10F4) (15D4). Cells were cultivated in a medium with 10 % FCS for three days, and subsequently in serum-free medium for two days. The procedure was repeated once again providing monoclonal antibody in the serum-free supernatants comparable in amounts and activity with that in serum containing media. The method enabled purification of antibodies by a single step in high purity.

Keywords: Hybridoma technology, monoclonal antibody, 1F5 , 10F4, 15D4, ELISA, serum-free medium, gel filtration chromotography

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