Department of Bioengineering, Guangdong Province Key Laboratory of Molecular Immunology and Antibody Engineering, Jinan University, Guangzhou 510632, PR China.
Department of Laboratory Medicine, Guangdong No. 2 Provincial People's Hospital, Guangzhou 510317, PR China.
Biosens Bioelectron. 2015 Jul 15;69:128-34. doi: 10.1016/j.bios.2015.02.026. Epub 2015 Feb 17.
Ultrasensitive and quantitative detection using simple and low-cost assays is critical in clinical diagnostics. In this report, we developed a triangular silver nanoprism (AgNPRs) etching-based plasmonic biosensor for the detection of cancer biomarkers. The triangular AgNPRs-based plasmonic biosensor is an enzyme-linked immunosorbent assay combined with the enzyme-mediated surface plasmon resonance (SPR) of triangular AgNPRs. Triangular AgNPRs uses the immune response of prostate-specific antigen (PSA) to trigger the glucose oxidase (GOx)-catalysed oxidation of glucose (Glu), producing hydrogen peroxide. Hydrogen peroxide acts as an oxidant to etch the triangular AgNPRs into smaller spherical silver nanoparticles, which is accompanied by a substantial blueshift of the SPR peak and a colourimetric blue-to-purple change that can be observed by the naked eye. The SPR peak shift enables the quantitative assessment of PSA due to the remarkable colour change. The triangular AgNPRs-based plasmonic ELISA approach exhibited a quasilinear response to logarithmic PSA concentrations in the range of 10fg/mL to 100pg/mL with a limit of detection (LOD) of 4.1fg/mL. In addition, the LOD of PSA in this approach exceeds that of the conventional HRP-based ELISA (1.25ng/mL) approach by more than 5 orders of magnitude. Patient serum samples from 16 donors were assayed with triangular AgNPRs-based plasmonic ELISA. The results from the triangular AgNPRs-based immunoassay and the time-resolved fluorescence immunoassay showed excellent correlation, and there were no significant differences in the quantified amounts of PSA. The triangular AgNPRs-based plasmonic ELISA approach has advantages (ultrasensitive, cost-effective, ease of operation) that are expected to be of great interest in diagnostics and to be suitable for a point-of-care test.
超灵敏和定量检测对于临床诊断至关重要。在本报告中,我们开发了一种基于三角形银纳米棱镜(AgNPRs)刻蚀的等离子体生物传感器,用于检测癌症生物标志物。基于三角形 AgNPRs 的等离子体生物传感器是一种酶联免疫吸附测定法,结合了酶介导的三角形 AgNPRs 的表面等离子体共振(SPR)。三角形 AgNPRs 利用前列腺特异性抗原(PSA)的免疫反应触发葡萄糖氧化酶(GOx)催化葡萄糖(Glu)的氧化,产生过氧化氢。过氧化氢作为氧化剂将三角形 AgNPRs 刻蚀成更小的球形银纳米粒子,同时伴随着 SPR 峰的显著蓝移和肉眼可见的比色蓝紫色变化。由于显著的颜色变化,SPR 峰的移动可以实现对 PSA 的定量评估。基于三角形 AgNPRs 的等离子体酶联免疫吸附测定法(ELISA)对 PSA 的对数浓度在 10fg/mL 至 100pg/mL 的范围内表现出准线性响应,检测限(LOD)为 4.1fg/mL。此外,与传统的基于辣根过氧化物酶(HRP)的 ELISA(1.25ng/mL)方法相比,该方法的 PSA 的 LOD 超过 5 个数量级。对来自 16 个供体的患者血清样本进行了基于三角形 AgNPRs 的等离子体 ELISA 检测。基于三角形 AgNPRs 的免疫测定和时间分辨荧光免疫测定的结果显示出极好的相关性,并且在 PSA 的定量量方面没有显著差异。基于三角形 AgNPRs 的等离子体 ELISA 方法具有超灵敏、经济高效、操作简便等优点,有望在诊断中引起极大兴趣,并适合于即时检测。