Wu Hong, Huo Qisheng, Varnum Susan, Wang Jun, Liu Guodong, Nie Zimin, Liu Jun, Lin Yuehe
Pacific Northwest National Laboratory, Richland, WA 99352, USA.
Analyst. 2008 Nov;133(11):1550-5. doi: 10.1039/b719810h. Epub 2008 Jul 28.
We report a dye-encapsulated silica nanoparticle as a label, with the advantages of high fluorescence intensity, photostability, and biocompatibility, in conjunction with microarray technology for sensitive immunoassay of a biomarker, interleukin-6 (IL-6), on a microarray format. The tris(2,2'-bipyridyl)ruthenium(ii) chloride hexahydrate (Rubpy) dye was incorporated into silica nanoparticles using a simple one-step microemulsion synthesis. In this synthesis process, Igepal CA520 was used as the surfactant, therefore, no requirement of cosolvent during the synthesis and the particle size was reduced comparing to the commonly used Triton surfactant system. The nanoparticles are uniform in size with a diameter of 50 nm. The microarray fluorescent immunoassay approach based on dye-doped silica nanoparticle labels has high sensitivity for practical applications with a limit of detection for IL-6 down to 0.1 ng mL(-1). The calibration curve is linear over the range from 0.1 ng mL(-1) to 10 ng mL(-1). Furthermore, results illustrated that the assay is highly specific for IL-6 in the presence of range of cytokines or proteins. The RuDS dye-labeled nanoparticles in connection with protein microarrays show the promise for clinical diagnosis of biomarkers.
我们报道了一种染料包封的二氧化硅纳米颗粒作为标记物,其具有高荧光强度、光稳定性和生物相容性等优点,并结合微阵列技术,以微阵列形式对生物标志物白细胞介素-6(IL-6)进行灵敏的免疫测定。采用简单的一步微乳液合成法将六水合三(2,2'-联吡啶)钌(II)氯化物(Rubpy)染料掺入二氧化硅纳米颗粒中。在该合成过程中,使用Igepal CA520作为表面活性剂,因此,合成过程中不需要助溶剂,并且与常用的Triton表面活性剂体系相比,粒径减小。纳米颗粒尺寸均匀,直径为50nm。基于染料掺杂二氧化硅纳米颗粒标记物的微阵列荧光免疫测定方法在实际应用中具有高灵敏度,IL-6的检测限低至0.1 ng mL(-1)。校准曲线在0.1 ng mL(-1)至10 ng mL(-1)范围内呈线性。此外,结果表明,在多种细胞因子或蛋白质存在的情况下,该测定对IL-6具有高度特异性。与蛋白质微阵列相关的RuDS染料标记纳米颗粒显示出用于生物标志物临床诊断的前景。