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基于生物素-链霉亲和素偶联的鲁棒免疫传感系统,用于空间定位的飞克毫升级白细胞介素-6 的检测。

Robust immunosensing system based on biotin-streptavidin coupling for spatially localized femtogram mL level detection of interleukin-6.

机构信息

ARC Centre of Excellence for Nanoscale Biophotonics (CNBP), Macquarie University, North Ryde 2109, Australia.

ARC Centre of Excellence for Nanoscale Biophotonics (CNBP), Macquarie University, North Ryde 2109, Australia; Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, PR China.

出版信息

Biosens Bioelectron. 2018 Apr 15;102:80-86. doi: 10.1016/j.bios.2017.11.023. Epub 2017 Nov 6.

DOI:10.1016/j.bios.2017.11.023
PMID:29127899
Abstract

Detection of a very low amount of cytokines such as interleukin-6 (IL-6) in clinical fluids is important in biomedical research and clinical applications. Here, we demonstrate spatially-localized ultrasensitive (femtogram mL) level detection of IL-6 in serum and in cell culture media. Our approach is based on a sandwich immunosensor fabricated on the surface of an optical fibre. Firstly, the biotinylated IL-6 capture antibody was immobilized on the fibre surface by biotin-streptavidin coupling. Then the fabricated fibre was used for capturing IL-6 followed by exposure to detection antibody which was labeled with the fluorescent magnetic nanoparticles to report the signal. A linear relationship between IL-6 concentration and the fluorescence signal was obtained in the range from 0.4pgmL to 400pgmL of IL-6, with the limit of detection down to 0.1pgmL. In addition, this optical fibre sensor was successfully applied for the localized detection of IL-6 with the spatial resolution of 200µm and a sample volume of 1µL. Finally, the performance of the fibre sensor was demonstrated by detection of IL-6 secreted by BV-2 cells with comparable performance of the conventional enzyme-linked immunosorbent assay (ELISA).

摘要

在生物医学研究和临床应用中,检测临床液体中诸如白细胞介素-6(IL-6)等非常低量的细胞因子非常重要。在这里,我们展示了在血清和细胞培养介质中对 IL-6 进行空间定位的超灵敏(皮克毫升级)检测。我们的方法基于在光纤表面制造的夹心免疫传感器。首先,通过生物素-链霉亲和素偶联将生物素化的 IL-6 捕获抗体固定在纤维表面上。然后,将制造的光纤用于捕获 IL-6,然后暴露于用荧光磁性纳米颗粒标记的检测抗体以报告信号。在 IL-6 浓度为 0.4pgmL 至 400pgmL 的范围内,IL-6 浓度与荧光信号之间呈线性关系,检测限低至 0.1pgmL。此外,该光纤传感器还成功地用于以 200µm 的空间分辨率和 1µL 的样品体积对 IL-6 进行局部检测。最后,通过检测 BV-2 细胞分泌的 IL-6 证明了纤维传感器的性能,其性能可与传统的酶联免疫吸附测定(ELISA)相媲美。

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