依托咪酯单克隆抗体的制备与应用

Preparation and Application of Monoclonal Antibody against Etomidate

目的:制备抗依托咪酯(ET)抗体,应用于胶体金方法学的ET快速检测试剂。方法:通过化学方法在ET分子苯环上引入氨基并连接上载体蛋白,合成ET人工抗原,经质谱和紫外光谱鉴定后,免疫Balb/C小鼠。采用杂交瘤技术得到高特异性ET杂交瘤细胞株。经Protein G过柱纯化,选择灵敏度合适的抗体应用于ET胶体金试剂上,通过药物交叉干扰实验及临床试验对其进行敏感性与特异性鉴定。结果:ET人工抗原合成成功,其免疫所得10F9细胞株抗体适用于ET胶体金免疫层析试剂。检定尿液和烟油ET最低检出限均为500 ng/mL,检测12种烟油添加物及48种常用药物未有交叉反应,临床试验得出其敏感性为100%,特异性为99.5%。结论:成功制备抗依托咪酯单克隆抗体(mAb)并应用于胶体金免疫层析试剂上。

Objective:To prepare anti-ET antibody for rapid detection of ET using the colloidal gold method.Methods:After identified by mass spectrometry and ultraviolet spectrum,the synthetic ET antigen was used to immunize Balb/C mice.ET hybridoma cell lines with high specificity were obtained by hybridoma technique.After purification by Protein G column,the antibody with appropriate sensitivity was selected and applied to ET colloidal gold reagent.The sensitivity and specificity of the antibody were identified by drug cross interference experiment and clinical experiment.Results:The antibody produced by 10F9 cell line was suitable for ET colloidal gold immunochromatography reagent.The ET reagent was used to detect 12 kinds of e-liquid additives and 48 kinds of commonly used drugs without cross-reaction.The cutoff of ET reagent in urine and e-liquid were both 500 ng/mL,the sensitivity was 100%,and the specificity was 99.5%.Conclusions:The monoclonal antibody(mAb)against ET was successfully prepared and applied to the colloidal gold immunochromatographic reagent.