title: [Preparation and characterization of the monoclonal antibodies against human CA19-9 and establishment of a sandwich CLIA system].
abstract: OBJECTIVE
  To prepare the monoclonal antibodies (mAbs) against human carbohydrate antigen 19-9 (CA19-9) and establish a double-antibody sandwich chemiluminescent immunoassay (CLIA) detection system for CA19-9 in the human serum.
  
  
  METHODS
  BALB/c mice were immunized with human CA19-9 antigen. The mAbs were obtained by hybridoma technique. The purity, titer, specificity and pairing of the mAbs were characterized and the sandwich CLIA system was established. The system was evaluated in its accuracy, limit of detection, linearity and repetitiveness after optimization of coating buffer, coating concentration and pipetting mode, and the serum sample was tested with it.
  
  
  RESULTS
  Four mAbs named #1-1, #2-1, #3-1 and #4-1 were obtained against human CA19-9. The titers of the anti-CA19-9 mAbs were above 10(-8). The mAbs had nearly no cross-reaction with CA125, CA15-3 and CA72-4. The double-antibody sandwich CLIA system was established by #3-1 mAb and #2-1 mAb-HRP. After optimization, its property was detection range of 0-1 000 U/mL, limit detection of 2.0 U/mL, linear correlation coefficient of 0.9999. The results which were contrasted with Roche test showed that: Kappa>0.75, r(correlation coefficient)>0.9.
  
  
  CONCLUSIONS
  The mAbs against human CA9-9 have been prepared and a sandwich CLIA system for detecting CA19-9 in the human serum has been established successfully.
authors.affiliation: College of Life Science, Northeast Forestry University, Harbin 150040, China.
authors.forename: Xingguo
authors.initials: X
authors.lastname: Lan
authors.affiliation: College of Life Science, Northeast Forestry University, Harbin 150040; Daqing MabCom, Inc., Daqing 163316, China.
authors.forename: Linggai
authors.initials: L
authors.lastname: Cao
authors.affiliation: College of Life Science, Northeast Forestry University, Harbin 150040; Daqing MabCom, Inc., Daqing 163316, China.
authors.forename: Deqiang
authors.initials: D
authors.lastname: Wei
authors.affiliation: Daqing MabCom, Inc., Daqing 163316, China.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Zhao
authors.affiliation: College of Life Science, Northeast Forestry University, Harbin 150040; 3Daqing 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*; D018395:CA-19-9 Antigen / Q000276:immunology*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D018395:CA-19-9 Antigen