[Preparation and characterization of monoclonal antibodies against human CA125 and establishment of a chemiluminescence immunoassay system].

OBJECTIVE:To prepare the anti-human carbohydrate antigen 125 (CA125) monoclonal antibody (mAb), and establish a chemiluminescence immunoassay detection system. METHODS:The BALB/c mice were immunized with human CA125 antigen. The hybridoma cells were obtained by cell fusion and screening. The mAbs were purified using protein A. The specificity, purity, isotype, and affinity of the mAbs were characterized and the sandwich ELISA system was established. The chemiluminescence immunoassay system was established and evaluated after optimization and selection of coating buffer, coating concentration, pipetting mode respectively, and the blood samples were tested with it. RESULTS:Three hybridoma cell lines named 30-1-4, 31-1-5 and 43-1-6 were obtained respectively. Anti-CA125 mAbs nearly had no cross-reaction with CA199, CA50 and CA724. The titres were above 1:10(8);. The chemiluminescence immunoassay system was established using 30-1-4 and 31-1-5. After optimization, the linear detection of the system covered a range 0-1000 U/mL, and its lowest detectable limit was 0.72 U/mL. The correlation coefficient with the results of the Roche test was above 0.90. CONCLUSION:With the mAbs against human CA125 we prepared, a chemiluminescence immunoassay system for human CA125 detection has been established successfully, which provides a basis for human CA125 detection and ovarian cancer diagnosis.