title: [Preparation and characterization of monoclonal antibodies against human CA125 and establishment of a chemiluminescence immunoassay system].
abstract: 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.
  
  
  CONCLUSIONS
  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.
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: College of Life Sciences, Northeast Forestry University, Harbin 150040, China.
authors.forename: Xingguo
authors.initials: X
authors.lastname: Lan
authors.affiliation: Daqing MabCom. Inc, Daqing 163316, China.
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Zhang
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 / Q000276:immunology*; D018394:CA-125 Antigen / Q000097:blood / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010051:Ovarian Neoplasms / Q000175:diagnosis
chemical_list: D000911:Antibodies, Monoclonal; D018394:CA-125 Antigen