title: [Preparation and characterization of monoclonal antibodies against human CA50 and establishment of chemiluminescence immunoassay system].
abstract: OBJECTIVE
  To prepare the anti-human carbohydrate antigen 50(CA50) monoclonal antibody (mAb), characterize its immunological features and establish a chemiluminescence immunoassay system with it.
  
  
  METHODS
  BALB/c mice were immunized with human CA50 antigen for preparing mAb using hybridoma technique. Stable anti-CA50-secreting hybridoma cell lines were obtained after screening. The mAbs 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 1:10(8). Anti-CA50 mAbs 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/mL. The recovery rate for accuracy was 107.08% and its sensitivity was 0.83 U/mL. The assay was highly repeatable [coefficient of variation (CV)<10%]. The correlation coefficient with the test of reference reagent was 0.96.
  
  
  CONCLUSIONS
  The monoclonal antibodies against human CA50 had been screened and a chemiluminescence immunoassay system for human CA50 detection had been established successfully.
authors.affiliation: College of Life Sciences, Northeast Forestry University, Harbin 150040; Daqing MabCom. Inc, Daqing 163316; Daqing Biotechnology Research Institute, Northeast Forestry University, Daqing 163316, China.
authors.forename: Xingguo
authors.initials: X
authors.lastname: Lan
authors.affiliation: College of Life Sciences, Northeast Forestry University, Harbin 150040; Daqing MabCom. Inc, Daqing 163316, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Daqing MabCom. Inc, Daqing 163316; Daqing Biotechnology Research Institute, Northeast Forestry University, Daqing 163316, China.
authors.forename: Deqiang
authors.initials: D
authors.lastname: Wei
authors.affiliation: College of Life Sciences, Northeast Forestry University, Harbin 150040; Daqing Biotechnology Research Institute, Northeast Forestry University, Daqing 163316, China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000276:immunology; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000032:analysis / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D049449:Luminescence; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; C082699:CA-50 antigen