Multiplexed electrochemical immunoassay for two immunoglobulin proteins based on Cd and Cu nanocrystals,Analyst

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Multiplexed electrochemical immunoassay for two immunoglobulin proteins based on Cd and Cu nanocrystals
Analyst ( IF 3.6 ) Pub Date : 2017-11-08 00:00:00 , DOI: 10.1039/c7an01459g
Dianping Tang _{1,

2,

3,

4,

5} , Jingjing Ren _{4,

5,

6,

7,

8} , Minghua Lu _{4,

9,

10,

11,

12}

Affiliation

Herein, a simple and feasible electrochemical immunosensing method for simultaneous voltammetric detection of two immunoglobulin proteins, human IgG (HIgG) and rabbit IgG (RIgG), was developed using two distinguishable signal-generation tags on the same electrode. The immunosensor was prepared by immobilizing two Fab antibody fragments on a gold electrode. After this, Cu and Cd nanocrystals, as nanotags, were synthesized and functionalized with identical detection antibodies. Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR) were employed to characterize the Cu and Cd nanocrystals. The covalently modified electrode with the Fab antibody fragments through the Au-thiolate bond (to dispel the non-specific adsorption) was investigated via scanning electron microscopy (SEM). After the sandwiched immunoreaction, the antibody-modified nanocrystals were captured on the immunosensor, which could be interrogated in pH 3.5 HCl using square-wave anodic stripping voltammetry. Experimental results also indicated that the multiplexed immunoassay enabled the simultaneous detection of HIgG and RIgG in a single run with the similar linear range from 0.01 to 10 ng mL⁻¹, and the limits of detection (LODs) towards two analytes could be as low as 3.4 pg mL⁻¹ (at 3σ). Acceptable assay results on precision, reproducibility, specificity, and method accuracy were also acquired. Importantly, the newly designed strategy avoided cross-talk and enzymatic introduction as compared to conventional electrochemical immunoassays, thus exhibiting a promising potential in clinical applications.

中文翻译：

基于Cd和Cu纳米晶体的两种免疫球蛋白蛋白质的多重电化学免疫测定

本文中，在同一电极上使用两个可区分的信号产生标签，开发了一种简单可行的电化学免疫传感方法，用于同时伏安法检测两种免疫球蛋白蛋白，人IgG（HIgG）和兔IgG（RIgG）。通过将两个Fab抗体片段固定在金电极上来制备免疫传感器。此后，合成了Cu和Cd纳米晶体作为纳米标签，并使用相同的检测抗体进行了功能化。透射电子显微镜（TEM）和傅立叶变换红外光谱（FTIR）用于表征Cu和Cd纳米晶体。通过所述的Au-硫醇盐键的Fab抗体片段的共价修饰的电极（消除非特异性吸附）进行了研究通过扫描电子显微镜（SEM）。夹在中间的免疫反应后，抗体修饰的纳米晶体被捕获在免疫传感器上，可以使用方波阳极溶出伏安法在pH 3.5 HCl中进行询问。实验结果还表明，多重免疫分析能够在0.01至10 ng mL ^-1的线性范围内一次检测HIgG和RIgG ，并且对两种分析物的检测限（LOD）可能低至3.4 pg毫升^-1（在3σ）。还获得了关于精密度，重现性，特异性和方法准确性的可接受的测定结果。重要的是，与常规的电化学免疫测定相比，新设计的策略避免了串扰和酶的引入，因此在临床应用中显示出有希望的潜力。

更新日期：2017-11-21

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