Tsai Pei-I, Lee Adam Shih-Yuan, Lee Shu-Sheng, Chung Ming-Han, Liu Meng-Wei, Lee Chih-Kung
Department of Engineering Science & Ocean Engineering, National Taiwan University, Taipei, Taiwan.
Department of Chemistry, Tamkang University, New Taipei City, Taiwan.
PLoS One. 2016 Jul 26;11(7):e0160031. doi: 10.1371/journal.pone.0160031. eCollection 2016.
For rapid screening and quantification of an antisera antibody, a nanometer bithiophene-based conductive biolinker can enhanced signal performance and can be used to verify the interaction of an anti-IFN-γ antibody with an IFN-γ protein. The experimental measurements take a generic approach which takes advantage of the functionality of thiophene-based linkers for biosensors. Effects associated with using bithiophene as a biolinker for surface plasmon resonance (SPR) spectroscopy are examined in this paper. By using an atomic force microscope (AFM), it was observed that the morphology of the bithiophene modified gold sensor surface became smoother than the original gold surface. We compared the response and concentration of the anti-IFN-γ antibody on a bithiophene-coated and dextran-coated biochip as well as on different thickness-modified surfaces under SPR relevant conditions. The results indicate that a response to IFN-γ molecules immobilized on a sensor using a bithiophene biolinker improved more than 8-fold when compared to that of a sensor using a dextran biolinker. Furthermore, the regeneration ability of the sensor surface shows good repeatability as only less than a 1% decrease was found after repeating the experimental work over 6 cycles. The characteristics provided us with a good platform for rapid screening, real-time monitoring and quantitative concentration of the autoimmune antibody activities.
对于抗血清抗体的快速筛选和定量,基于纳米双噻吩的导电生物连接体可增强信号性能,并可用于验证抗IFN-γ抗体与IFN-γ蛋白的相互作用。实验测量采用一种通用方法,该方法利用了基于噻吩的连接体对生物传感器的功能。本文研究了使用双噻吩作为表面等离子体共振(SPR)光谱的生物连接体所产生的影响。通过使用原子力显微镜(AFM)观察到,双噻吩修饰的金传感器表面的形态比原始金表面更光滑。我们比较了在SPR相关条件下,抗IFN-γ抗体在双噻吩涂层和葡聚糖涂层生物芯片以及不同厚度修饰表面上的响应和浓度。结果表明,与使用葡聚糖生物连接体的传感器相比,使用双噻吩生物连接体固定在传感器上的IFN-γ分子的响应提高了8倍以上。此外,传感器表面的再生能力显示出良好的重复性,在重复实验工作6个循环后,发现下降幅度仅小于1%。这些特性为自身免疫抗体活性的快速筛选、实时监测和定量浓度提供了一个良好的平台。