Production of Monoclonal Antibodies Specific for Progesterone, Estradiole by Simultaneous Injection of Different Steroids


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.

Keywords: Immunodiagnosis, hybrid cell, monoclonal antibody, steroid hormones, ELISA.

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