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.
ABSTRACT
Objective To construct and identify norepinephrine ( NE) complete antigen for the preparation of high sensitive and specific anti-NE monoclonal antibody .Methods Glutaraldehyde ( GA) and 1-Ethyl-3-(3-Dimethylaminopropyl ) carbodiimide ( EDC) were used to cross-link NE with carrier pro-teins (BSA, OVA) for NE complete antigen preparation under conditions of pH 4.5 or pH9.0.Three assays including UV scanning , SDS-PAGE and FeCl3 color reaction were performed for identification of NE com-plete antigen.Serum antibody titers were evaluated in mice model induced by intraperitoneal immunization with NE complete antigen .Results NE complete antigens were successfully prepared as indicated by the three identification assays .The coupling ratio was significantly increased in a time-depended manner under the condition of pH9.0 in comparison to that in the condition of pH 4.5.Indirect ELISA results showed that , when coating antigens and serum antibodies were prepared with the same cross -linking method , the serum antibody titers were significantly higher than those with different methods .Conclusion Anti-NE antibodies were successfully prepared by immunizing mice with NE complete antigens .