Preparation against carbohydrate antigen 19-9 monoclonal antibodies and estab-lishment of DAS-ELISA
10.3969/j.issn.1000-484X.2014.08.018
- VernacularTitle:糖类抗原19-9单抗制备及DAS-ELISA的建立
- Author:
Yunlong WANG
;
Jinyu ZHAI
;
Jichuang WANG
;
Lei CHENG
;
Yulin LI
;
Xinjie GE
;
Lie MAO
- Publication Type:Journal Article
- Keywords:
CA19-9;
Monoclonal antibody;
DAS-ELISA
- From:
Chinese Journal of Immunology
2014;(8):1088-1092
- CountryChina
- Language:Chinese
-
Abstract:
To prepare monoclonal antibody of carbohydrate antigen 19-9(CA19-9).Methods: Based on the titer test results of mouse ascites and its IC 50 values ,the mouse that prepare for fusion was identified.Positive monoclonal cell strains were established by cell fusing and screening.Monoclonal antibody from ascites was produced by peritoneal injection monoclonal cell , and then purified by octoic acid-ammonium sulfate precipitation method.After determine the protein concentrations by UV-spectrophotometry ,the monoclonal antibody against CA 19-9 was labelled with horseradish peroxidase.Based on antibody pairing test , DAS-ELISA method was established .To compared with abroad kit , analyzing performance of this method.Results: Three strains of monoclonal antibody were obtained.And the optimal working concentrations of mAb (ZJY3-1G9) ,as coated antibody,McAb(ZJY2-7F10),as HRP-IgG,were assured.Limit of detection was 26.4 U/ml.Linear range was 30-300 U/ml.By detecting patients with serum 33 , confirmed the correlation coefficient of r=0.950 4 , compared with abroad kit that measure simultaneously.Conclusion:Monoclonal antibody prepared for CA 19-9 can be used to develop a kit.
