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用于检测血清中人类白细胞介素-8的无标记电化学阻抗生物传感器,灵敏度可达亚皮克/毫升。

Label-free electrochemical impedance biosensor to detect human interleukin-8 in serum with sub-pg/ml sensitivity.

作者信息

Sharma R, Deacon S E, Nowak D, George S E, Szymonik M P, Tang A A S, Tomlinson D C, Davies A G, McPherson M J, Wälti C

机构信息

School of Electronic and Electrical Engineering, University of Leeds, Woodhouse Lane, Leeds LS2 9JT, UK.

School of Molecular and Cellular Biology, University of Leeds, Woodhouse Lane, Leeds LS2 9JT, UK.

出版信息

Biosens Bioelectron. 2016 Jun 15;80:607-613. doi: 10.1016/j.bios.2016.02.028. Epub 2016 Feb 11.

Abstract

Biosensors with high sensitivity and short time-to-result that are capable of detecting biomarkers in body fluids such as serum are an important prerequisite for early diagnostics in modern healthcare provision. Here, we report the development of an electrochemical impedance-based sensor for the detection in serum of human interleukin-8 (IL-8), a pro-angiogenic chemokine implicated in a wide range of inflammatory diseases. The sensor employs a small and robust synthetic non-antibody capture protein based on a cystatin scaffold that displays high affinity for human IL-8 with a KD of 35 ± 10 nM and excellent ligand specificity. The change in the phase of the electrochemical impedance from the serum baseline, ∆θ(ƒ), measured at 0.1 Hz, was used as the measure for quantifying IL-8 concentration in the fluid. Optimal sensor signal was observed after 15 min incubation, and the sensor exhibited a linear response versus logarithm of IL-8 concentration from 900 fg/ml to 900 ng/ml. A detection limit of around 90 fg/ml, which is significantly lower than the basal clinical levels of 5-10 pg/ml, was observed. Our results are significant for the development of point-of-care and early diagnostics where high sensitivity and short time-to-results are essential.

摘要

具有高灵敏度和短检测时间、能够检测血清等体液中生物标志物的生物传感器是现代医疗保健早期诊断的重要前提。在此,我们报告了一种基于电化学阻抗的传感器的开发,用于检测血清中的人白细胞介素-8(IL-8),这是一种促血管生成的趋化因子,与多种炎症性疾病有关。该传感器采用了一种基于半胱氨酸蛋白酶抑制剂支架的小型且坚固的合成非抗体捕获蛋白,其对人IL-8具有高亲和力,解离常数为35±10 nM,且具有出色的配体特异性。在0.1 Hz下测量的血清基线的电化学阻抗相位变化∆θ(ƒ)被用作定量流体中IL-8浓度的指标。孵育15分钟后观察到最佳传感器信号,并且该传感器对IL-8浓度的对数在900 fg/ml至900 ng/ml范围内呈现线性响应。观察到检测限约为90 fg/ml,明显低于5-10 pg/ml的基础临床水平。我们的结果对于即时检测和早期诊断的发展具有重要意义,其中高灵敏度和短检测时间至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b92/4785862/421a8c328c0a/gr1.jpg

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