抗人CA19-9单克隆抗体的制备和鉴定及化学发光检测体系的建立

发布时间：2018-11-14 11:16

【摘要】：目的制备抗人糖抗原19-9(CA19-9)单克隆抗体(mAb),建立双抗体夹心检测CA19-9化学发光体系。方法 CA19-9免疫小鼠后,通过细胞融合技术,获得抗CA19-9的mAb。鉴定其抗体的纯度、效价、特异性和配对,建立双抗体夹心化学发光体系。在包被缓冲液、包被浓度及加样模式上优化化学发光体系,对该体系进行准确度、最低检测限、线性、重复性评估以及血清样本的检测。结果获得4株名称为#1-1、#2-1、#3-1和#4-1的mAb。抗体效价均大于10-8,与CA125、CA15-3、CA72-4均无交叉反应。其中#3-1 mAb与#2-1-HRP建立的双抗体夹心体系经优化后,其检测范围为0~1 000 U/mL;最低检测限为2.0 U/mL;线性相关系数为0.9999。与罗氏试剂检测结果对比,Kappa系数大于0.75;相关系数大于0.9。结论成功制备了抗人CA19-9 mAb,建立了双抗体夹心检测CA19-9化学发光体系。
[Abstract]:Aim to prepare anti-human glucose antigen 19-9 (CA19-9) monoclonal antibody (mAb), and to establish a double antibody sandwich system for the detection of CA19-9 chemiluminescence. Methods after immunizing mice with CA19-9, mAb. against CA19-9 was obtained by cell fusion technique. The purity, titer, specificity and pairing of the antibody were identified and a double antibody sandwich chemiluminescence system was established. The chemiluminescence system was optimized in the buffer solution, the coating concentration and the sample mode. The accuracy, the minimum detection limit, the linearity, the repeatability and the detection of serum samples were evaluated. Results four mAb. strains named # 1-1 #2-1 #3-1 and # 4-1 were obtained. The antibody titers were higher than 10-8, and there was no cross reaction with CA125,CA15-3,CA72-4. The detection range of the double antibody sandwich system established by # 3-1 mAb and # 2-1-HRP is 0 ~ 1 000 U / mL, the lowest detection limit is 2.0 U / mL, and the linear correlation coefficient is 0.9999 9. Compared with the results of Roche reagent, the Kappa coefficient was greater than 0.75 and the correlation coefficient was greater than 0.9. Conclusion the double antibody sandwich system for the detection of CA19-9 chemiluminescence was successfully prepared by anti-human CA19-9 mAb,.
【作者单位】：东北林业大学生命科学学院;大庆麦伯康生物技术有限公司;东北林业大学大庆生物技术研究院;
【基金】：黑龙江省应用技术研究与开发项目(GC13C122) 黑龙江省发展高新技术产业专项资金(FW11C201)
【分类号】：R392.1

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