Rapid detection of bladder cancer using an immunoassay transistor combined with DNA-labeling technique in a microfluidic chip

Po-Yu Peng, W. Hsu, T. Pan, Yen-Heng Lin
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引用次数: 1

Abstract

We propose a method that can be used to quantitatively measure concentrations of protein through a semiconductor sensor with a robust signal. Measurement of protein by a semiconductor may encounter a major problem. For specifically measuring protein, an antibody is immobilized on the semiconductor sensor surface. The physical length of the antibody is around 10 nm, a distance too long to be sensed by the semiconductor sensor, which means that the following attached protein cannot be sensed. In this study, we used a bead-based immunoassay combined with the DNA strain labeling technique to overcome this issue. Protein was first captured using an antibody-coated magnetic bead. It was then labeled with a secondary antibody combined with the DNA strain. Finally, the magnetic bead with the biotarget was attracted on the sensor surface by an external magnetic field, and then the negative charges of the DNA changed the surface potential of the sensor. The concentration of protein could then be measured accordingly. The signal could be further improved by the optimization of the number of labeling DNA and the size of the magnetic bead. In addition, the semiconductor sensor was incorporated in a microfluidic chip with microvalves and a micromixer. The microfluidic procedure reduced the total measurement time to around 1 h (plate ELISA 4h). Furthermore, by changing the antibody to another type of antibody coated on the magnetic bead, the sensor could be reused to measure other types of protein. The proposed method provides a solution for the robust measurement of protein concentration through the semiconductor sensor.
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微流控芯片中结合dna标记技术的免疫分析晶体管快速检测膀胱癌
我们提出了一种方法,可用于定量测量浓度的蛋白质通过半导体传感器与鲁棒信号。用半导体测量蛋白质可能会遇到一个大问题。为了特异性地测量蛋白质,在半导体传感器表面固定了抗体。抗体的物理长度约为10nm,这个距离太长,半导体传感器无法感知,这意味着下面附着的蛋白质无法被感知。在这项研究中,我们使用了基于头部的免疫分析结合DNA菌株标记技术来克服这个问题。首先用抗体包覆的磁珠捕获蛋白质。然后用二抗结合DNA菌株进行标记。最后,在外加磁场的作用下,带有生物靶点的磁珠被吸引到传感器表面,DNA的负电荷改变了传感器的表面电位。然后就可以测量蛋白质的浓度。通过优化标记DNA的数量和磁珠的大小,可以进一步提高信号的质量。此外,还将半导体传感器集成在带有微阀和微混合器的微流控芯片中。微流控程序将总测量时间缩短至约1小时(平板ELISA为4小时)。此外,通过将抗体改变为涂在磁珠上的另一种类型的抗体,传感器可以重复使用来测量其他类型的蛋白质。该方法为通过半导体传感器实现蛋白质浓度的鲁棒测量提供了一种解决方案。
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