硼酸化量子点荧光微球免疫层析试纸条定量检测甲胎蛋白  

Boronic acid-functionalized quantum dot nanobeads basedimmunochromatographic strip for quantitative detection of alpha fetoprotein

作　　者：林童 郭徐静 武敬宇 熊勇华[1] 郭亮[1] LIN Tong;GUO Xujing;WU Jingyu;XIONG Yonghua;GUO Liang(Jiangxi-OAI Joint Research Institute,Nanchang University,Nanchang 330047,China)

机构地区：[1]南昌大学中德联合研究院,江西南昌330047

出　　处：《南昌大学学报（理科版）》2021年第4期358-364,共7页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(31360385);国家重点研发计划项目(2018YFC1602505)。

摘　　要：提高表面抗体活性是免疫检测探针制备的关键。硼酸可与IgG的糖基化Fc片段快速发生位点特异性结合,充分暴露抗原结合区Fab片段以提高抗体活性,避免常用的酰胺键共价偶联方法引起的抗体取向不当及自交联。本研究在合成硼酸修饰的聚马来酸酐-alt-1-十八碳烯(PBA-PMAO)的基础上,采用超声乳化法制备表面硼酸改性的量子点荧光微球(PBA-QBs)。该微球既可与抗体简便、快速地定向偶联,又因量子点含量高而具有很强的荧光信号。将其与甲胎蛋白(AFP)单克隆抗体室温共孵育5 min制备高抗体活性的荧光免疫层析检测探针并用于定量检测AFP,线性范围为0.98~31.25 ng mL^(-1),最低检测限为0.45 ng mL^(-1),检测耗时20 min。血清样品中AFP的加标回收率为91.0%~107.1%,批内、批间变异系数介于3.65%~10.42%。Improving the activities of the antibodies on the surface is crucial to the preparation of immunoprobe.Boric acid can conjugate to the glycosylated Fc fragment of IgG quickly.Therefore,the antigen-binding region,namely Fab fragment,will be exposed to facilitate the antibody activity via the site-specific conjugation,avoiding the improper antibody oriention and antibody crosslink introduced by conventional amide-conjugation.In this study,phenylboronic acid-functionalized quantum dot nanobeads(PBA-QBs)were prepared via a facile mini-emulsion by using the as-synthesised phenylboronic acid modified poly(maleic anhydride-alt-1-octadecene)(PBA-PMAO)and quantum dots.The PBA-QBs were capable of conjugating to antibodies orientedly and emitting strong fluorescence ascribed to the boronic moieties on its surface and the numerous encapsulated quantum dots,respectively.Then the fluorescent immunoprobes with high antibody activity for immunochromatographic assay(ICA)of alpha fetoprotein(AFP)were prepared via a 5 min-incubation of PBA-QBs and anti-AFP antibodies at room temperature.The PBA-QBs-based ICA took only 20 min,and displayed a dynamic linear detection of 0.49~15.6 ng mL^(-1) with a detection limit of 0.46 ng mL^(-1).Furthermore,PBA-QBs-based immunoprobe was used to analyze the AFP-spiked serum with recoveries ranging from 91.0%to 107.1%and intra-and inter-assay coefficients of variation ranging from 3.65%to 10.42%.

关 键 词：免疫检测探针 硼酸 量子点荧光微球 甲胎蛋白 

分 类 号：O65[理学—分析化学]