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Production and application of monoclonal antibody specific for human chorionic gonadotropin
M Stamenova1, I Kehayov, S Kyurkchiev
1Department of Biology, Bulgarian Medical Academy, Sofia.
Spleen cells from mice immunized with human chorionic gonadotropin (hCG) were fused with mouse myeloma cells and four hybridomas, secreting monoclonal antibodies, were selected for further studies. In cross-check tests against tissue extracts and peptide hormones it was established that mAb IB10 (IgG1) reacted positively against hCG only but not with other hormones. Ascites from this hybridoma was fractionated by ammonium sulphate precipitation and by FPLC to isolate the IgG fraction. Subsequently, the purified mAb was conjugated with horseradish peroxidase and used for development of a simple test for pregnancy diagnosis. Some preliminary experiments have shown that the test is very specific and reproducible.
Spleen cells from mice immunized with human chorionic gonadotropin (hCG) were fused with mouse myeloma cells and four hybridomas, secreting monoclonal antibodies, were selected for further studies. In cross-check tests against tissue extracts and peptide hormones it was established that mAb IB10 (IgG1) reacted positively against hCG only but not with other hormones. Ascites from this hybridoma was fractionated by ammonium sulphate precipitation and by FPLC to isolate the IgG fraction. Subsequently, the purified mAb was conjugated with horseradish peroxidase and used for development of a simple test for pregnancy diagnosis. Some preliminary experiments have shown that the test is very specific and reproducible.