该研究旨在制备具有高磁、光学强度的 "磁包金"核壳异质型磁/金纳米材料(magnetic gold nanoparticles,MGNPs),并将其作为标记探针构建高灵敏检测猪尿中盐酸克伦特罗(clenbuterol,Clen)的免疫层析新方法。使用乳液自组装法合成MGNPs,采用EDC法在其表面偶联Clen单克隆抗体制备双功能信号探针,建立基于MGNPs试纸条定量检测猪尿中Clen的竞争抑制曲线,以确定其线性范围和灵敏度。采用加标回收实验评价试纸条的准确性和精密度,使用商业化的ELISA试剂盒评价试纸条的实用性和可靠性。结果表明,所合成的MGNPs具有更高的磁饱和强度(为初始氧化铁纳米粒子的62.2%)和更好的光学信号强度(为传统30 nm胶体金纳米粒子的8倍)。基于MGNPs的试纸条定量检测猪尿中Clen的线性范围为0.078~40 ng/mL,半数抑制浓度为1.225 ng/mL,最低检出限为0.041 ng/mL,检测时间为15 min。加标回收实验显示,试纸条批内、批间回收率为93.66% ~ 114.80%,变异系数小于13.45%。此外该方法与商业化ELISA试剂盒检测结果具有良好的相关性。
This study aims to prepare a novel core-shell heterostructure magnetic gold nanoparticles (MGNPs) with high magnetic and optical strength,and then it was used as a bifunctional probe to establish a new immunochromatographic method (ICA) for highly-sensitive detection of clenbuterol (Clen) in swine urine.A facile emulsion self-assembly strategy was used to synthesize MGNPs,and the EDC method was used to prepare a bifunctional signal probe by coupling anti-Clen monoclonal antibody onto the surface of MGNPs.Moreover,the competition inhibition curve for the quantitative detection of Clen was established,and the dynamic detection range and detection sensitivity of the developed MGNP-ICA were then determined.The accuracy and precision of the MGNPs-ICA were also been evaluated by a standard recovery experiment,and the reliability of the MGNPs-ICA was estimated by a commercial ELISA kit.Results showed that the as-prepared MGNPs have high saturation magnetization (62.2% of the initial iron oxide nanoparticles) and strong absorbance (8 times higher than that of traditional 30 nm colloidal gold nanoparticles).Taking advantage of their excellent magnetic properties and optical sensing capacity,the detection range of MGNPs-ICA for Clen was 0.078-40 ng/mL.Furthermore,the half-maxinal inhibitory concentration was 1.225 ng/mL with the limit of detection 0.041 ng/mL.And this assay could be finished in 15 min.The recoveries of intra- and inter-assay of MGNPs-ICA range from 93.66% to 114.80% with the coefficient of variation (CV) of below 13.45%.In addition,this method had a good correlation with the detection results obtained from commercial ELISA kits.
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