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《Chinese Journal of Cellular and Molecular Immunology》 2015-01
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Preparation and characterization of monoclonal antibodies against human CA50 and establishment of chemiluminescence immunoassay system

LAN Xingguo;FENG Yue;WEI Deqiang;LI Yuhua;College of Life Sciences,Northeast Forestry University;Daqing Mab Com.Inc;Daqing Biotechnology Research Institute,Northeast Forestry University;  
Objective To prepare the anti-human carbohydrate antigen 50(CA50) monoclonal antibody(m Ab),characterize its immunological features and establish a chemiluminescence immunoassay system with it.Methods BALB/c mice were immunized with human CA50 antigen for preparing m Ab using hybridoma technique.Stable anti-CA50-secreting hybridoma cell lines were obtained after screening.The m Abs were purified using protein A after expanding culture.The chemiluminescence immunoassay system was established and evaluated in its linear range,accuracy,sensitivity,reproducibility,and the blood samples were tested with it.Results Four hybridoma cell lines were obtained respectively and their titers were above 1∶108.Anti-CA50 m Abs nearly had no cross-reaction with CA125,CA153,CA199 and CA724.The linear detection of the chemiluminescence immunoassay system covered a range 0-500 U/m L.The recovery rate for accuracy was 107.08% and its sensitivity was 0.83 U/m L.The assay was highly repeatable [coefficient of variation(CV) 10%].The correlation coefficient with the test of reference reagent was 0.96.Conclusion The monoclonal antibodies against human CA50 had been screened and a chemiluminescence immunoassay system for human CA50 detection had been established successfully.
【Fund】: 黑龙江省应用技术研究与开发项目(GC13C122);; 黑龙江省发展高新技术产业专项资金(FW11C201)
【CateGory Index】: R392
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