Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden.
Global Research and Development, Thermo Fisher Scientific IDD, Uppsala, Sweden.
Sci Rep. 2017 Jul 28;7(1):6837. doi: 10.1038/s41598-017-07030-0.
The introduction of nanomaterials as detection reagents has enabled improved sensitivity and facilitated detection in a variety of bioanalytical assays. However, high nanoprobe densities are typically needed for colorimetric detection and to circumvent this limitation several enhancement protocols have been reported. Nevertheless, there is currently a lack of universal, enzyme-free and versatile methods that can be readily applied to existing as well as new biosensing strategies. The novel method presented here is shown to enhance the signal of gold nanoparticles enabling visual detection of a spot containing <10 nanoparticles. Detection of Protein G on paper arrays was improved by a 100-fold amplification factor in under five minutes of assay time, using IgG-labelled gold, silver, silica and iron oxide nanoprobes. Furthermore, we show that the presented protocol can be applied to a commercial allergen microarray assay, ImmunoCAP ISAC sIgE 112, attaining a good agreement with fluorescent detection when analysing human clinical samples.
纳米材料作为检测试剂的引入提高了灵敏度,并促进了各种生物分析检测中的检测。然而,对于比色检测通常需要高的纳米探针密度,为了规避这一限制,已经报道了几种增强方案。然而,目前缺乏通用的、无酶的和多功能的方法,可以很容易地应用于现有的和新的生物传感策略。这里提出的新方法被证明可以增强金纳米粒子的信号,从而实现对包含<10个纳米粒子的点的可视化检测。使用 IgG 标记的金、银、硅和氧化铁纳米探针,在不到五分钟的检测时间内,将免疫球蛋白 G 固定在纸阵列上的检测提高了 100 倍的放大倍数。此外,我们表明,所提出的方案可以应用于商业过敏原微阵列检测,ImmunoCAP ISAC sIgE 112,当分析人类临床样本时,与荧光检测具有良好的一致性。