tailieunhanh - Simple production and purification of monoclonal antibodies in serum free medium

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. | Turk J Biol 24 (2000) 189–196 © TÜBİTAK Simple Production and Purification of Monoclonal Antibodies in Serum-Free Medium Aynur BAŞALP Institute for Genetic Engineering and Biotechnology, TUBITAK, Marmara Research Center, 41470 Gebze, Kocaeli-TURKEY Beyazıt ÇIRAKOĞLU Department of Medical Biology, Medical Faculty, Marmara University, 81326 Haydarpaşa, Istanbul-TURKEY Engin BERMEK Department of Biophysics, Istanbul Medical Faculty, 34390 Çapa, Istanbul-TURKEY Received: 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 17β-estradiol (10F4) (15D4). Cells were cultivated in a medium with 10 % FCS for three days, and subsequently in serumfree 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. Key Words: Hybridoma technology, monoclonal antibody, 1F5 , 10F4, 15D4, ELISA, serum-free medium, gel filtration chromotography Monoklonal Antikorların Basit Bir Yöntemle Serumsuz Besi Yerinde Üretilmesi ve Saflaştırılması Özet: Serum hücreler için çok gerekli temel besi maddelerini ve büyüme faktörlerini içermesine rağmen, serum kullanımı besi ortamının bileşiminin kompleks ve tanımlanamaz hale gelmesine neden olur. Özellikle antikorların saflaştırılması ve karakterizasyonunda, serumsuz besi ortamı serumlu besi yerinden daha etkilidir. Bu çalışmada Tütün Mozaik Virüsü (1F5) ve 17β-östradiol’e karşı geliştirilmiş olan monoklonal .