摘要
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 macrophage inhibitory cytokine-1 （MIC-1） monoclonal antibody and the test method for serum MIC-1. Methods BALB/C female mice was immuned by MIC-1 antigen. Then hybridoma cell lines secreting monoclonal antibodies against MIC-1 were obtained by hybridoma technology. ELISA serum test method was con- structed and performance was evaluated. Results Totally 32 monoclonal antibodies against MIC-1 were successfully ob- tained. Two hihg-affinity antiboclies were chosen to establish MIC-1 serum rapid detection method. Performance appraisal results showed good linearity, and R2 is greater than 0. 999. Intra-and inter-assay coefficient of variation were 5.15% and 9. 51% respectively. The average recovery was 98.9%. The MIC-ldetection method was stable at 37℃ for up to 3 days and 4℃ for 6 months. Conclusion The indicators of the MIC-1 detection method conform to the related state standards or industrial standards. The MIC-1 ELISA serum test method meets the need of scientific research and clinical test and has entered the industrialization program.
出处
《癌症进展》
2011年第4期361-366,共6页
Oncology Progress
基金
国家高技术研究发展863计划资助项目(项目编号:2008AA02Z415)
