不同类型探针对免疫层析方法的分析灵敏度影响研究  

Study on Influences of Different Probes on Analytical Sensitivity of Immunochromatography Assay

作　　者：马嘉琪 姚晓琳 刘思杰 殷雪驰 毕鹏飞 王建龙[1] 张道宏[1] MA Jia-qi;YAO Xiao-lin;LIU Si-jie;YIN Xue-chi;BI Peng-fei;WANG Jian-long;ZHANG Dao-hong(College of Food Science and Engineering,Northwest Agriculture and Forestry University,Yangling 712100,China)

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100

出　　处：《分析测试学报》2021年第3期310-317,共8页Journal of Instrumental Analysis

基　　金：陕西省重点研发计划项目(2019NY-109);大学生创新创业训练计划项目(X202010712305)。

摘　　要：为研究不同类型探针对免疫层析方法的分析灵敏度影响,该研究以17β-雌二醇(17β-E2)为检测对象,采用磁性纳米材料标记7μg/mL的17β-E2单克隆抗体(检测抗体)制备了传统探针,采用磁性纳米材料分别标记5μg/mL的17β-E2单克隆抗体和5μg/mL的羊抗鼠IgG多克隆抗体(第二抗体)制备了配对探针,采用磁性纳米材料标记14μg/mL的羊抗鼠IgG多克隆抗体制备了间接探针。然后,基于这3种不同类型的探针,分别建立了17β-E2的免疫层析快速检测方法,并对3种17β-E2免疫层析方法的检测灵敏度进行比较。结果显示,基于传统、配对和间接探针的免疫层析方法对17β-E2的检出限分别为1、0.5、0.2 ng/mL,其中基于间接探针的免疫层析方法灵敏度最高,与基于传统探针和配对探针的免疫层析方法相比,检出限分别降低了5倍和2.5倍。该研究刷新了传统免疫层析方法中探针必须是信号材料标记检测抗体的认知,可为其他食品安全危害因子的免疫层析快速检测研究提供有价值的参考。To study the influences of different probes on the analytical sensitivity of immunochromatography assay(ICA),a traditional probe was prepared for the detection of 17β-estradiol(17β-E2),with magnetic nanoparticles(MNPs)to label 7μg/mL 17β-E2 monoclonal antibody(mAb,detection Ab).Meanwhile,a paired probe was prepared by using MNPs to separately label 5μg/mL 17β-E2 mAb and 5μg/mL goat anti-mouse polyclonal antibody(pAb,the second antibody),and an indirect probe was prepared by using MNPs to label 14μg/mL pAb.Then,three immunochromatography assays were established based on the three probes for the rapid detection of estradiol.The analytical sensitivities of three ICAs were compared by the 17β-E2 detection.Results showed that the analytical sensitivities of ICAs based on traditional,paired and indirect probes for 17β-E2 are 1,0.5,0.2 ng/mL,respectively.Among these three ICAs,the analytical sensitivity of indirect probe based ICA is the highest.Compared with the traditional and paired probes based ICAs,the detection limit of the indirect probe based ICA is 5 fold and 2.5 fold reduced,respectively.This study refreshes the conventional cognition toward the signal probe which is considered can only be prepared by using signal materials to label the detection Ab in the traditional immunochromatography assay,and provides a valuable reference for the rapid detection of other food safety hazard factors by immunochromatography assays.

关 键 词：竞争免疫层析 探针 17β-雌二醇(17β-E2) 食品安全 

分 类 号：O658.1[理学—分析化学]