基于金磁微粒与酶标噬菌体抗体的磁分离免疫分析法

Magnetic Affinity Immunoassay Based on Gold-magnetic Particle and HRP-conjugated Phage Antibody

基于金磁微粒（Gold-magnetic particle）兼有纳米金颗粒与磁微粒特性的优势,以相思子毒素（Abrin）为目标物,将蛋白A（SPA）包被金磁微粒偶联多抗作为功能化捕获探针,酶标噬菌体抗体作为特异信号检测探针,建立了一种检测相思子毒素的磁分离免疫分析法。该方法的线性范围为0.008~250μg/L,相关系数（r）为0.991 0,检出限为0.008μg/L,定量下限为0.008μg/L。该方法将蛋白A-金磁微粒功能化探针与酶标噬菌体抗体探针的优势结合,提高了检测灵敏度、特异性和抗干扰能力,适用于各种环境样品中微量相思子毒素样品的分析。

A new magnetic affinity immunoassay（ MAIA） method based on gold-magnetic particle and HRP-conjugated phage antibody was developed. Gold-magnetic particle were coated with staphylococcus protein A（ SPA） and coupled with polyclonal antibody（ pc Ab） to construct functionalized capturing probes,and HRP-conjugated phage antibody was then used as specific detecting probe to design MAIA for trace detection of abrin. The calibration curve was linear（ r = 0. 991 0） in the range of0. 008- 250 μg / L,with a limit of detection（ LOD） of 0. 008 μg / L, and a limit of quantitation（ LOQ） of 0. 008 μg /L. Integrating the unique advantages of functionalized SPA-gold-magnetic particle probe and HRP-conjugated phage antibody with its high sensitivity,specificity and anti-interference,the method is proven to be well suited for the analysis of trace abrin in various environmental samples.