A disposable electrochemical immunosensor for the determination of leptin in serum and breast milk
A disposable electrochemical immunosensor for the determination of leptin in serum and breast milk
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DOI：
10.1039/c3an00183k
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发表时间：
2013-01-01
期刊：
影响因子：
4.2
通讯作者：
Pingarron, Jose M.
中科院分区：
文献类型：
--
作者：
Ojeda, Irene;Moreno-Guzman, Maria;Pingarron, Jose M.
本文介绍了一种用于定量测定瘦素的一次性电化学免疫传感器的制备方法。的制备方法涉及固定化的一个特定的生物素化的抗瘦素抗体的表面上的链霉亲和素功能化的磁珠（Strept-MBs）和一个ESTA型免疫测定涉及的目标分析物，单克隆抗瘦素，和IgG标记的碱性磷酸酶（AP-IgG）。电化学转导步骤通过借助于Nd磁体将携带免疫缀合物的MB捕获到丝网印刷碳电极（SPCE）上并通过差分脉冲伏安法（DPV）测量在加入1-萘基磷酸（1-NPP）后AP酶反应中产生的1-萘酚的电化学氧化来完成。线性范围为5 ~ 100 pg mL（-1），检出限为0.5 pg mL（-1）（3sb/m）。该值比迄今为止文献中描述的唯一的瘦素伏安免疫传感器中报道的值低27倍以上。通过分析不同类型的真实的样品证明了免疫传感器的有用性：人血清、婴儿奶粉和来自母乳喂养两个月的哺乳母亲的母乳。
The preparation of a disposable electrochemical immunosensor for the quantification of the hormone leptin is described in this work. The preparation approach involved immobilization of a specific biotinylated anti-leptin antibody on the surface of streptavidin-functionalized magnetic beads (Strept-MBs) and a sandwich-type immunoassay involving the target analyte, monoclonal anti-leptin, and IgG labeled with alkaline phosphatase (AP-IgG). The electrochemical transduction step was accomplished by trapping the MBs bearing the immunoconjugates onto screen-printed carbon electrodes (SPCEs) by means of an Nd magnet and measuring the electrochemical oxidation of the 1-naphthol generated in the AP enzyme reaction upon 1-naphthyl phosphate (1-NPP) additions by differential pulse voltammetry (DPV). A calibration plot with a linear range between 5 and 100 pg mL(-1) as well as a detection limit of 0.5 pg mL(-1) (3sb/m) were achieved. This value is more than 27 times lower than that reported in the only voltammetric immunosensor for leptin described in the literature until now. The usefulness of the immunosensor was demonstrated by analyzing different types of real samples: human serum, infant powdered milk, and breast milk from a nursing mother with two months of breastfeeding.