CAJ | 학술논문

目的：为制备糖类抗原19-9（Carbohydrate antigen 19-9，CA19-9）特异性单克隆抗体。方法：以免疫后小鼠血清效价及其IC50值确定融合用小鼠，融合、筛选获得阳性杂交瘤细胞株。制备、收集并纯化腹水抗体，紫外分光光度法测定抗体浓度；辣根过氧化物酶标记抗体后，筛选配对抗体建立DAS-ELISA检测方法，并与国外检测试剂盒进行对比验证检测性能。结果：细胞融合后获得3株单抗细胞株（ZJY1-2C8、ZJY2-7F10、ZJY3-1G9），筛选确定ZJY3-1G9作为包被抗体、ZJY2-7F10作为酶标抗体，成功建立DAS-ELISA检测方法，最低检测限为26.4 U/ml。线性范围为30～300 U/ml。通过检测病患血清33份，证实与国外试剂盒检测的相关系数r＝0.9504。结论：所制备的单抗可用于CA19-9试剂盒的研制。
목적：위제비당류항원19-9（Carbohydrate antigen 19-9，CA19-9）특이성단극륭항체。방법：이면역후소서혈청효개급기IC50치학정융합용소서，융합、사선획득양성잡교류세포주。제비、수집병순화복수항체，자외분광광도법측정항체농도；랄근과양화물매표기항체후，사선배대항체건립DAS-ELISA검측방법，병여국외검측시제합진행대비험증검측성능。결과：세포융합후획득3주단항세포주（ZJY1-2C8、ZJY2-7F10、ZJY3-1G9），사선학정ZJY3-1G9작위포피항체、ZJY2-7F10작위매표항체，성공건립DAS-ELISA검측방법，최저검측한위26.4 U/ml。선성범위위30～300 U/ml。통과검측병환혈청33빈，증실여국외시제합검측적상관계수r＝0.9504。결론：소제비적단항가용우CA19-9시제합적연제。
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.