Phurimsak Chayakom, Tarn Mark D, Pamme Nicole
Department of Chemistry, University of Hull, Cottingham Road, Hull, HU6 7RX, UK.
Micromachines (Basel). 2016 Apr 26;7(5):77. doi: 10.3390/mi7050077.
Conventional immunoassays offer selective and quantitative detection of a number of biomarkers, but are laborious and time-consuming. Magnetic particle-based assays allow easy and rapid selection of analytes, but still suffer from the requirement of tedious multiple reaction and washing steps. Here, we demonstrate the trapping of functionalised magnetic particles within a microchannel for performing rapid immunoassays by flushing consecutive reagent and washing solutions over the trapped particle plug. Three main studies were performed to investigate the potential of the platform for quantitative analysis of biomarkers: (i) a streptavidin-biotin binding assay; (ii) a sandwich assay of the inflammation biomarker, C-reactive protein (CRP); and (iii) detection of the steroid hormone, progesterone (P4), towards a competitive assay. Quantitative analysis with low limits of detection was demonstrated with streptavidin-biotin, while the CRP and P4 assays exhibited the ability to detect clinically relevant analytes, and all assays were completed in only 15 min. These preliminary results show the great potential of the platform for performing rapid, low volume magnetic particle plug-based assays of a range of clinical biomarkers via an exceedingly simple technique.
传统免疫测定法能够选择性地定量检测多种生物标志物，但操作繁琐且耗时。基于磁性颗粒的测定法能轻松快速地筛选分析物，但仍需进行繁琐的多重反应和洗涤步骤。在此，我们展示了在微通道内捕获功能化磁性颗粒，通过向捕获的颗粒塞冲洗连续的试剂和洗涤溶液来进行快速免疫测定。进行了三项主要研究以探究该平台对生物标志物进行定量分析的潜力：（i）链霉亲和素-生物素结合测定；（ii）炎症生物标志物C反应蛋白（CRP）的夹心测定；（iii）针对竞争性测定的类固醇激素孕酮（P4）的检测。链霉亲和素-生物素实现了低检测限的定量分析，而CRP和P4测定显示出检测临床相关分析物的能力，并且所有测定仅需15分钟即可完成。这些初步结果表明，该平台具有巨大潜力，可通过极其简单的技术对一系列临床生物标志物进行基于磁性颗粒塞的快速、微量测定。
