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目的研制巨噬细胞抑制因子-1(MIC-1)单克隆抗体并建立MIC-1血清检测方法。方法应用自主研制的MIC-1抗原免疫BALB/C雌鼠,通过杂交瘤技术获得分泌抗MIC-1单克隆抗体的杂交瘤细胞株,通过ELISA方法构建MIC-1血清检测体系并对性能进行鉴定。结果成功获得了32株抗MIC-1单克隆抗体,选用其中2株高亲和力单克隆抗体建立了双夹心ELISA MIC-1血清快速检测方法。性能鉴定结果表明所建立的方法线性关系良好(R2值大于0.999);试验内变异系数为5.15%,试验间变异系数为9.51%;平均回收率为98.9%;37℃保存3天和4℃保存6个月稳定性良好。结论所制备的MIC-1检测方法各项指标均达到SFDA相关要求,能够满足科学研究和临床检测的技术需要,并进入产业化程序。
Objective To develop a monoclonal antibody against macrophage inhibitory factor-1 (MIC-1) and to establish a method for the detection of serum MIC-1. Methods BALB / C female mice were immunized with self-developed MIC-1 antigen, hybridoma cell lines secreting anti-MIC-1 monoclonal antibody were obtained by hybridoma technique, and the MIC-1 serum detection system was constructed by ELISA and its performance was identified . Results 32 anti-MIC-1 monoclonal antibodies were successfully obtained and 2 high-affinity monoclonal antibodies were selected to establish a rapid sandwich ELISA-MIC-1 serum test. The results of performance appraisal showed that the established method had a good linear relationship (R2 value> 0.999); the coefficient of variation (CV) was 5.15% and the coefficient of variation (CV) was 9.51%; the average recovery was 98.9% 6 months stability is good. Conclusion All the indexes of MIC-1 detection method meet the requirements of SFDA, meet the technical requirements of scientific research and clinical testing, and enter the industrialization process.