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Nanopore blockade sensors for ultrasensitive detection of proteins in complex biological samples.
Nature Communications ( IF 14.7 ) Pub Date : 2019-05-08 , DOI: 10.1038/s41467-019-10147-7
Kyloon Chuah 1 , Yanfang Wu 1 , S R C Vivekchand 1 , Katharina Gaus 2 , Peter J Reece 3 , Adam P Micolich 3 , J Justin Gooding 1
Affiliation  

Nanopore sensors detect individual species passing through a nanoscale pore. This experimental paradigm suffers from long analysis times at low analyte concentration and non-specific signals in complex media. These limit effectiveness of nanopore sensors for quantitative analysis. Here, we address these challenges using antibody-modified magnetic nanoparticles ((anti-PSA)-MNPs) that diffuse at zero magnetic field to capture the analyte, prostate-specific antigen (PSA). The (anti-PSA)-MNPs are magnetically driven to block an array of nanopores rather than translocate through the nanopore. Specificity is obtained by modifying nanopores with anti-PSA antibodies such that PSA molecules captured by (anti-PSA)-MNPs form an immunosandwich in the nanopore. Reversing the magnetic field removes (anti-PSA)-MNPs that have not captured PSA, limiting non-specific effects. The combined features allow detecting PSA in whole blood with a 0.8 fM detection limit. Our 'magnetic nanoparticle, nanopore blockade' concept points towards a strategy to improving nanopore biosensors for quantitative analysis of various protein and nucleic acid species.

中文翻译:

用于对复杂生物样品中的蛋白质进行超灵敏检测的纳米孔阻滞传感器。

纳米孔传感器检测通过纳米孔的单个物种。这种实验范式在低分析物浓度和复杂介质中的非特异性信号下存在较长的分析时间。这些限制了纳米孔传感器用于定量分析的有效性。在这里,我们使用在零磁场下扩散的抗体修饰的磁性纳米粒子((抗 PSA)-MNPs)来解决这些挑战,以捕获分析物、前列腺特异性抗原 (PSA)。(anti-PSA)-MNPs 被磁力驱动以阻塞纳米孔阵列,而不是通过纳米孔移位。通过用抗 PSA 抗体修饰纳米孔,使(抗 PSA)-MNP 捕获的 PSA 分子在纳米孔中形成免疫夹心,从而获得特异性。反转磁场可去除未捕获 PSA 的(抗 PSA)-MNP,限制非特异性作用。这些组合功能允许检测全血中的 PSA,检测限为 0.8 fM。我们的“磁性纳米颗粒、纳米孔封锁”概念指向了一种改进纳米孔生物传感器以对各种蛋白质和核酸种类进行定量分析的策略。
更新日期:2019-05-16
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