Ultrasensitive multiplexed immunoassay of autophagic biomarkers based on Au/rGO and Au nanocages amplifying electrochemcial signal.
Ultrasensitive multiplexed immunoassay of autophagic biomarkers based on Au/rGO and Au nanocages amplifying electrochemcial signal.
复制标题
基于 Au/rGO 和 Au 纳米笼放大电化学信号的自噬生物标志物的超灵敏多重免疫分析
DOI：
10.1038/s41598-017-02766-1
复制
发表时间：
2017-05-26
影响因子：
4.6
通讯作者：
Mei X
中科院分区：
文献类型：
--
作者：
Wang G;Li Y;Liu J;Yuan Y;Shen Z;Mei X
首次提出了一种新的自噬生物标志物的电化学免疫传感器，金还原氧化石墨烯纳米复合物（Au/r-GO）作为一个良好的导电平台，具有超高的比表面积，为Beclin-1和LC 3B-II两种抗体提供了更多的结合位点。而Au纳米笼（AuNCs）由于其表面含有丰富的活性氧官能团，可以作为良好的导电平台，封装大量的氧化还原探针和二抗，用于信号放大。通过差分脉冲伏安法（DPV）测量，可以在单次运行中直接检测到两个独立的信号，这代表贝林-1和LC 3B-II的存在。在优化的条件下，Beclin-1和LC 3B-II的线性范围为0.1 ~ 100 ng/mL，具有良好的灵敏度和选择性。Beclin-1和LC 3B-II的检测限分别为0.02和0.03 ng/mL。将该方法应用于实验细胞蛋白裂解液中Beclin-1和LC 3B-II的检测，结果与酶联免疫吸附试验结果一致。该方法为自噬的检测提供了一种简便、灵敏、定量的方法
A novel sandwich-assay electrochemical immunosensor for simultaneous determination of autophagic biomarkers was introduced for the first time, the gold-reduced grapheme oxide nanocomposite (Au/r-GO) set as a good conductive platform with super high specific area, and provided more binding sites for the both antibodies of Beclin-1 and LC3B-II. While Au nanocages (AuNCs) served as good conductive platform to encapsulate a large amount of redox probe and secondary antibodies for signal amplification, due to the abundant reactive oxygen functional groups on its surface. Through differential pulse voltammetry (DPV) measurements, two separate signals can be detected directly in a single run, which represent the existence of Belin-1 and LC3B-II. Under optimized conditions, the electrochemical immunosensor exhibited good sensitivity and selectivity for the simultaneous determination of Beclin-1 and LC3B-II with linear ranges of 0.1–100 ng/mL. The detection limit for Beclin-1 and LC3B-II is 0.02 and 0.03 ng/mL respectively. This method was also applied for the analysis of Beclin-1 and LC3B-II levels in experimental cellular protein lysates, and the results were in good agreement with those of enzyme linked immunosorbent assay. This approach gives a promising simple, sensitive and quantitative strategy for the detection of autophagy