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首页> 外文期刊>Bioprocess and Biosystems Engineering >Conductometric Immunoassay For Interleukin-6 In Human Serum Based On Organic/inorganic Hybrid Membrane-functionalized Interface
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Conductometric Immunoassay For Interleukin-6 In Human Serum Based On Organic/inorganic Hybrid Membrane-functionalized Interface

机译：基于有机/无机杂化膜功能化界面的人血清白细胞介素6电导免疫测定

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摘要

Various sensor-based immunoassay methods have been extensively developed for the detection of interleukin-6 (IL6), but most often exhibit low detection signals and low detection sensitivity, and are unsuitable for routine use. The aim of this work is to develop a simple and sensitive conductometric immunoassay for IL6 in human serum by using an organic/inorganic hybrid membrane-functionalized interface. Initially, thionine-bound 3,4,9,10-perylenetetracarboxylic acid was doped into colloidal alumina, then nanogold particles were immobilized onto the thionine surface, and then horseradish peroxidase-labeled anti-IL6 antibodies were conjugated on the nanogold surface. The organic/inorganic hybrid membrane provides a good microenvironment for the immobilization of biomolecules, enhanced the surface coverage of protein, and improved the sensitivity of the immunosensor. The performance and factors influencing the performance of the immunosensor were evaluated. The detection is based on the change in local conductivity before and after the antigen-antibody interaction in 0.02 M phosphate buffer solution (pH 6.8) containing 50 μM H_2O_2, 0.01 M KI and 0.15 M NaCl. Under optimal conditions, the proposed immunosensor exhibited a wide linear range from 25 to 400 pg/ml towards IL6 with a relatively low detection limit of 5 pg/ml (S/N = 3). The stability, reproducibility and precision of the immunosensor were acceptable. 37 serum specimens were assayed by the developed immunosensor and standard enzyme-linked immunosorbent assay, respectively, and the results obtained were almost consistent. More importantly, the detection methodology provides a promising approach for other proteins or biosecuritys.
机译：已经广泛开发了各种基于传感器的免疫分析方法来检测白介素6（IL6），但大多数情况下显示出低检测信号和低检测灵敏度，并且不适合常规使用。这项工作的目的是通过使用有机/无机杂化膜功能化界面为人血清中的IL6开发一种简单而灵敏的电导免疫测定方法。最初，将硫氨酸结合的3,4,9,10-per四甲酸掺杂到胶体氧化铝中，然后将纳米金颗粒固定在硫氨酸表面上，然后将辣根过氧化物酶标记的抗IL6抗体偶联到纳米金表面上。有机/无机杂化膜为固定生物分子提供了良好的微环境，增加了蛋白质的表面覆盖率，并提高了免疫传感器的灵敏度。评估了性能和影响免疫传感器性能的因素。该检测基于抗原抗体在含有50μMH_2O_2、0.01 M KI和0.15 M NaCl的0.02 M磷酸盐缓冲溶液（pH 6.8）中相互作用前后的局部电导率变化。在最佳条件下，拟议的免疫传感器对IL6的线性范围为25至400 pg / ml，相对较低的检测限为5 pg / ml（S / N = 3）。免疫传感器的稳定性，可重复性和精确度是可以接受的。通过开发的免疫传感器和标准酶联免疫吸附法分别检测了37个血清样品，获得的结果几乎一致。更重要的是，检测方法为其他蛋白质或生物安全性提供了一种有前途的方法。

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