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Production of monoclonal antibodies specific for progesterone, estradiole by simultaneous injection of different steroids

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We report here the development of hybrid cells producing monoclonal antibodies specific for two different steroid hormones with mixed immunization using hybridoma technology. BALB/c mice were immunized with a mixture of three steroid antigens: progesterone, estradiole and testosterone linked to bovine serum albumine. | Turk J Biol 24 (2000) 697–705 © TÜBİTAK Production of Monoclonal Antibodies Specific for Progesterone, Estradiole by Simultaneous Injection of Different Steroids Fatıma ŞAHİNGÖZ YÜCEL Gen Mühendisliği ve Biyoteknoloji Araştırma Enstitüsü, TÜBİTAK Marmara Araştırma Merkezi, Gebze-Kocaeli-TURKEY Beyazıt ÇIRAKOĞLU Marmara Üniversitesi Tıp Fakültesi Tıbbi Biyoloji ve Genetik A.B.D. Haydarpaşa, İstanbul-TURKEY Received: 28.01.1999 Abstract: We report here the development of hybrid cells producing monoclonal antibodies specific for two different steroid hormones with mixed immunization using hybridoma technology. BALB/c mice were immunized with a mixture of three steroid antigens: progesterone, estradiole and testosterone linked to bovine serum albumine. These mice were used for fusion. In the two fusion experiments, ELISA tests showed that among 645 wells only 2 hybrids reacted with progesterone (MAM 3C2, MAM 3E3) and one of them reacted specifically with estradiol (MAM 7H7). Three types of monoclonal antibodies were detected: IgG1, IgG2a and IgG2b and the affinities of these antibodies were found to be 10-8–10-9 M. Monoclonal antibodies did not cross react with the other steroid hormones. Key Words: Immunodiagnosis, hybrid cell, monoclonal antibody, steroid hormones, ELISA. Eş Zamanlı İmmünizasyonla Progesteron Estradiol ve Testosteron’a Özgü Monoklonal Antikorların Birlikte Üretilmesi Özet: Bu makalede karma immünizasyon tekniği ve hibridoma teknolojisi kullanılarak iki farklı steroid hormona özgün monoklonal antikorlar sentezleyen hibrit hücrelerinin geliştirilmesi rapor edilmektedir. Her biri sığır serum albuminine bağlı progesteron, estradiol ve testosteron’dan oluşan steroid hormon karışımı ile BALB/c fareler immünize edilmiştir. Bu fareler füzyon için kullanılmıştır. İki füzyon deneyinde, 645 kuyuda hibrit klon gözlenmiş bu klonlar içerisinde 2 tanesinin (MAM 3C2, MAM 3E3) progesteron ile reaksiyon verdiği bir tanesinin de ( MAM 7H7) estradiole özgü olduğu .

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