Department of Chemistry, Loughborough University, Loughborough, Leicestershire, LE11 3TU, UK.
Analyst. 2020 Apr 7;145(7):2595-2601. doi: 10.1039/d0an00063a. Epub 2020 Feb 17.
Prion diseases are a group of fatal transmissible neurological conditions caused by the change in conformation of intrinsic cellular prion protein (PrP). We present a rapid assay using aptamers and resistive pulse sensing, RPS, to extract and quantify PrP from complex sample matrices. We functionalise the surface of superparamagnetic beads, SPBs, with a DNA aptamer. First SPB's termed P-beads, are used to pre-concentrate the analyte from a large sample volume. The PrP protein is then eluted from the P-beads before aptamer modified sensing beads, S-beads, are added. The velocity of the S-beads through the nanopore reveals the concentration of the PrP protein. The process is done in under an hour and allows the detection of picomol's of protein.
朊病毒病是一组由固有细胞朊病毒蛋白 (PrP) 构象改变引起的致命传染性神经疾病。我们提出了一种使用适体和电阻脉冲感应 (RPS) 的快速分析方法，从复杂的样品基质中提取和定量 PrP。我们用 DNA 适体功能化超顺磁性珠 (SPB) 的表面。首先，将 SPB 称为 P 珠，用于从大样品体积中预浓缩分析物。然后将 PrP 蛋白从 P 珠中洗脱出来，再加入经适体修饰的传感珠 (S 珠)。S 珠通过纳米孔的速度揭示了 PrP 蛋白的浓度。整个过程在一个小时内完成，可检测到皮摩尔级别的蛋白质。
