Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, People's Republic of China.
Talanta. 2013 Jan 15;103:392-7. doi: 10.1016/j.talanta.2012.10.087. Epub 2012 Nov 2.
This study developed a sensitive and selective aptamer-based chemiluminescent (CL) method for the determination of platelet-derived growth factor (PDGF)-BB using hydroxylamine enlarged gold nanoparticles (Au NPs). Rabbit anti-human PDGF-BB polyclonal antibody was covalently coupled on the 96-well plate that offers reactive N-oxysuccinimide ester (referred to as NOS group) surface. In the presence of target protein, the biotinylated aptamer was captured on the 96-well plate forming an antibody/PDGF-BB/biotinylated aptamer sandwiched complex, which was followed by the assembly of streptavidin coated Au NPs (streptavidin-gold). Au NPs assembled on the surface of 96-well plate reacted with HAuCl(4) and NH(2)OH, which enabled the catalytic deposition of gold metal onto the Au NPs surfaces. A huge number of Au(3+) ions were released from the hydroxylamine enlarged Au NPs after oxidative gold metal dissolution, which was determined by a simple and sensitive luminol CL reaction. The results showed that the detection limit of the assay is 60 pM of PDGF-BB (corresponding to 6 fmol in a 100 μL volume), which compares favorably with those of other PDGF-BB detection techniques. In addition, this aptameric CL biosensor demonstrated extraordinary specificity. And PDGF-BB has been determined in diluted serum indicating the applicability of this assay.
本研究开发了一种基于适体的灵敏和选择性化学发光（CL）方法，用于测定血小板衍生生长因子（PDGF-BB），该方法使用羟胺扩大的金纳米粒子（Au NPs）。兔抗人 PDGF-BB 多克隆抗体通过共价偶联到 96 孔板上，该孔板提供反应性 N-羟基琥珀酰亚胺酯（NOS 基团）表面。在存在靶蛋白的情况下，生物素化适体被捕获在 96 孔板上，形成抗体/PDGF-BB/生物素化适体夹心复合物，随后在金纳米粒子（streptavidin-gold）表面组装链霉亲和素。Au NPs 组装在 96 孔板表面上与 HAuCl(4)和 NH(2)OH 反应，这使得金金属能够在 Au NPs 表面上催化沉积。在金纳米粒子的氧化金金属溶解后，从羟胺扩大的 Au NPs 中释放出大量的 Au(3+)离子，这可以通过简单而灵敏的鲁米诺 CL 反应来确定。结果表明，该测定的检测限为 60 pM 的 PDGF-BB（相当于 100 μL 体积中的 6 fmol），这与其他 PDGF-BB 检测技术相比具有优势。此外，这种适体 CL 生物传感器表现出非凡的特异性。并且已经在稀释的血清中测定了 PDGF-BB，表明该测定法的适用性。
