Biotin-avidin-conjugated metal sulfide nanoclusters for simultaneous electrochemical immunoassay of tetracycline and chloramphenicol
Biotin-avidin-conjugated metal sulfide nanoclusters for simultaneous electrochemical immunoassay of tetracycline and chloramphenicol
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DOI：
10.1007/s00604-013-1096-2
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发表时间：
2014-01-01
影响因子：
5.7
通讯作者：
Tang, Dianping
中科院分区：
文献类型：
--
作者：
Liu, Bingqian;Zhang, Bing;Tang, Dianping
我们报告的协议，同时竞争性免疫测定四环素（TC）和氯霉素（CAP）在同一个传感接口。首先将TC和CAP与牛血清白蛋白的缀合物共固定在用纳米金修饰的玻碳电极上。平行地，单克隆抗TC和抗CAP抗体分别缀合到CdS和PbS纳米簇上。在典型的测定中，固定的半抗原和添加的靶分析物竞争结合纳米簇上的相应抗体。随后，镉（II）和铅（II）离子从相应的纳米团簇的表面释放的酸处理，然后通过方波阳极溶出伏安法检测。将Cd（II）和Pb（II）的峰值电位处的电流分别用作TC和CAP的传感器信号。这种多重免疫分析能够在单次运行中同时测定TC和CAP，两种分析物的动态范围为0.01至50 ng mL（-1）。TC和CAP的检测限分别为7.5 pg mL（-1）和5.4 pg mL（-1）。在一系列分析中未观察到明显的非特异性吸附和交叉反应性。批内和批间变异系数均小于10%。通过对牛奶和蜂蜜中TC和CAP的分析，对该方法进行了评价。回收率TC为88%~ 107%，CAP为91%~ 119%.
We report on a protocol for a simultaneous competitive immunoassay for tetracycline (TC) and chloramphenicol (CAP) on the same sensing interface. Conjugates of TC and of CAP with bovine serum albumin were first co-immobilized on a glassy carbon electrode modified with gold nanoparticles. In parallel, monoclonal anti-TC and anti-CAP antibodies were conjugated onto CdS and PbS nanoclusters, respectively. In a typical assay, the immobilized haptens and the added target analytes competed for binding to the corresponding antibodies on the nanoclusters. Subsequently, Cd(II) and Pb(II) ions are released from the surface of the corresponding nanoclusters by treatment with acid and then were detected by square wave anodic stripping voltammetry. The currents at the peak potentials for Cd(II) and Pb(II) were used as the sensor signal for TC and CAP, respectively. This multiplex immunoassay enables the simultaneous determination of TC and CAP in a single run with dynamic ranges from 0.01 to 50 ng mL(-1) for both analytes. The detection limits for TC and for CAP are 7.5 pg mL(-1) and 5.4 pg mL(-1), respectively. No obvious nonspecific adsorption and cross-reactivity was observed in a series of analyses. Intra-assay and inter-assay coefficients of variation were less than 10 %. The method was evaluated by analyzing TC and CAP in spiked samples of milk and honey. The recoveries range from 88 % to 107 % for TC, and from 91 % to 119 % for CAP.