Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, College of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, PR China.
Analyst. 2010 Aug;135(8):2036-42. doi: 10.1039/c0an00022a. Epub 2010 Jun 3.
A new strategy was described for amplifying the response of the antigen-antibody sensing processes by functionalizing SiO(2) nanoparticles labeled secondary antibodies based on a sandwich immunoassay in this work. At first, the multi-walled carbon nanotubes (CNTs) were individually dispersed in the aqueous bovine serum albumin (BSA) to obtain BSA molecules coated CNTs (BSA-CNTs). Then the gold colloids (nano-Au) were absorbed on the BSA-CNTs surface by the amido and disulfide groups of BSA. Later, a functionalized gold/carbon nanotube composite nanohybrid (DpAu/nano-Au/BSA-CNTs) modified electrode was developed by electrochemical deposition of Au(3+) onto nano-Au/BSA-MWNTs surface. Thus, a sensitive immunosensor for carbohydrate antigen 19-9 (CA19-9) has been constructed by further employment of Nafion coated SiO(2) nanoparticles labeled secondary antibody (SiO(2)-Ab(2)) for the signal amplification. More importantly, the loading of SiO(2)-Ab(2) can not only cause the construction of the dielectric antigen-antibody immunocomplex layer but also introduce the insulated Nafion coated SiO(2) nanoparticles which demonstrate the relatively high resistance, resulting in a strong detection signal. The proposed sensing strategy provides a wide linear dynamic range from 0.15 to 150 U mL(-1) with a low detection limit of 0.06 U mL(-1) at 3 times the background noise. Moreover, the extremely high stability of the functionalized gold/carbon nanotube composite nanohybrid monolayer allows the designed biosensing interface to obtain a good stability and long-term life.
本工作中描述了一种新策略,通过基于夹心免疫测定法对基于功能化 SiO(2)纳米粒子标记的二级抗体的 SiO(2)纳米粒子进行功能化,从而放大抗原-抗体传感过程的响应。首先,将多壁碳纳米管 (CNT) 单独分散在水性牛血清白蛋白 (BSA) 中,以获得涂覆 BSA 的 CNT(BSA-CNTs)分子。然后,通过 BSA 的酰胺和二硫键将金胶体 (纳米-Au) 吸附在 BSA-CNTs 表面上。之后,通过将 Au(3+)电化学沉积到纳米-Au/BSA-MWNTs 表面上,开发了一种功能化的金/碳纳米管复合纳米杂化(DpAu/nano-Au/BSA-CNTs)修饰电极。因此,通过进一步使用标记有二级抗体的涂覆有 Nafion 的 SiO(2)纳米粒子(SiO(2)-Ab(2))进行信号放大,构建了用于碳水化合物抗原 19-9(CA19-9)的灵敏免疫传感器。更重要的是,SiO(2)-Ab(2)的负载不仅可以引起介电抗原-抗体免疫复合物层的构建,而且还可以引入具有相对高电阻的涂覆有 Nafion 的 SiO(2)纳米粒子,从而产生强检测信号。所提出的传感策略提供了从 0.15 到 150 U mL(-1)的宽线性动态范围,检测下限为 0.06 U mL(-1),背景噪声为 3 倍。此外,功能化的金/碳纳米管复合纳米杂化单层的极高稳定性使设计的生物传感界面具有良好的稳定性和长期寿命。