论文部分内容阅读
目的制备抗氯胺酮(ketamine)的单克隆抗体(mAb),并鉴定其特性,为氯胺酮的胶体金免疫试纸条研制奠定基础。方法氯胺酮-BSA免疫BALB/c小鼠,采用杂交瘤技术制备抗氯胺酮的mAb。采用间接ELISA、竞争抑制ELISA和Western blot对mAb的特异性进行鉴定。采用间接ELISA法鉴定mAb的Ig亚类、检测其腹水效价及相对亲和力,并进行表位分析。结果获得2株可分泌特异性mAb的杂交瘤细胞(FB12b和CH10),其Ig亚类分别为IgG2a和IgG3;腹水mAb的效价均为1∶106;mAb FB12b和CH10的相对亲和力均在105以上。ELISA相加实验的结果显示,2株mAb可识别不同的抗原表位。结论成功制备抗氯胺酮的2株mAb,为建立快速特异检测氯胺酮滥用的现场检测方法提供了有力的工具。
Objective To prepare a monoclonal antibody (mAb) against ketamine and to identify its properties, which laid the foundation for the development of ketamine immunohistochemical strip for ketamine. Methods BALB / c mice were immunized with ketamine-BSA, and the anti-ketamine mAb was prepared by hybridoma technique. The specificity of mAb was identified by indirect ELISA, competitive inhibition ELISA and Western blot. The indirect ELISA was used to identify the Ig subclass of mAb. The ascites titer and relative affinity of the mAb were determined and the epitopes were analyzed. Results Two hybridoma cells (FB12b and CH10) secreting specific mAb were obtained. The Ig subclasses were IgG2a and IgG3, the titer of ascites mAb was 1:10, and the relative affinity of mAb FB12b and CH10 was 105 the above. The result of ELISA addition experiment shows that two mAbs can recognize different epitopes. Conclusion The successful preparation of 2 mAbs against ketamine provides a powerful tool to establish a rapid and specific detection of ketamine abuse in the field.