Selbes Yesim Somay, Caglayan Mehmet Gokhan, Eryilmaz Merve, Boyaci Ismail Hakki, Saglam Necdet, Basaran Arif Ahmet, Tamer Ugur
Turkish Doping Control Center, Hacettepe University, Sihhiye, 06100, Ankara, Turkey.
Department of Analytical Chemistry, Faculty of Pharmacy, Ankara University, 06100, Ankara, Turkey.
Anal Bioanal Chem. 2016 Nov;408(29):8447-8456. doi: 10.1007/s00216-016-9966-1. Epub 2016 Oct 8.
We present a surface-enhanced Raman probe (SERS) platform for the determination of a prohibited substance, recombinant erythropoietin (rEPO), in urine matrix, using nanoparticles as substrate. Rod-shaped gold nanoparticles (GNR) were modified with a Raman label and an antibody as SERS probe. We developed two SERS-based immunoassays for detection and quantification of rEPO in urine. In the first assay, rEPO was determined by a sandwich assay with gold surfaces and GNR. In the second assay, rEPO was extracted by using core shell-structured magnetic iron oxide gold nanoparticles, and again sandwich assay was performed by using GNR. We also demonstrated the ability of the proposed method to discriminate rEPO and urinary erythropoietin (uEPO). A good linear correlation was obtained between logarithms of rEPO concentrations in urine and Raman intensities within the range of 10-10 pg mL rEPO concentrations. Detection limits which are smaller than 0.1 pg mL levels were achieved owing to the high extractive performance of the nanoextraction techniques. Graphical Abstract Schematic represantation of surface-enhanced Raman probe for rapid nanoextraction and detection of erythropoietin.
我们展示了一种表面增强拉曼探针（SERS）平台，该平台以纳米颗粒为底物，用于测定尿液基质中的违禁物质重组促红细胞生成素（rEPO）。棒状金纳米颗粒（GNR）用拉曼标记物和抗体进行修饰，作为SERS探针。我们开发了两种基于SERS的免疫分析方法，用于检测和定量尿液中的rEPO。在第一种分析方法中，通过金表面和GNR的夹心分析来测定rEPO。在第二种分析方法中，使用核壳结构的磁性氧化铁金纳米颗粒提取rEPO，然后再次使用GNR进行夹心分析。我们还展示了所提出方法区分rEPO和尿促红细胞生成素（uEPO）的能力。在尿液中rEPO浓度为10 - 10 pg/mL的范围内，尿液中rEPO浓度的对数与拉曼强度之间获得了良好的线性相关性。由于纳米萃取技术的高萃取性能，实现了低于0.1 pg/mL水平的检测限。图形摘要 用于快速纳米萃取和检测促红细胞生成素的表面增强拉曼探针的示意图。
