【作者】 仇立群；

【导师】 顾仁敖；

【作者基本信息】 苏州大学 ， 物理化学， 2003， 硕士

【摘要】 表面增强拉曼光谱(SERS)技术作为最灵敏的检测界面物种的谱学技术之一，被越来越广泛的应用于各种分子水平的测试中，结合近红外傅立叶变换拉曼光谱仪，将其应用于标记免疫检测将有非常大的应用前景。本文试图通过免疫金标记技术将SERS引入标记免疫检测，主要进行了以下几方面的工作： 以苯硫酚为探针，讨论了金溶胶粒径与SERS强度的关系，通过金种生长法制得可在近红外傅立叶变换拉曼光谱仪上得到较好SERS信号的金溶胶，粒径约为50nm。同时，还发现组装金粒子在生长后其粒径增长也使SERS强度得到增强。 用上述生长金溶胶制备稳定的SERS标记免疫溶胶。主要探讨了制备过程中SERS标记分子苯硫酚合适的加入量，反应时间；抗体羊抗小鼠IgG与SERS标记溶胶的结合条件等。 由于我们设想以银片为免疫检测中固相抗体的基底，所以论文中首先以SERS标记的免疫金溶胶为探针，讨论了抗体与银表面间的作用，实验中发现，SERS标记的免疫金溶胶可以通过抗体与银之间的作用较好的自组装于银表面，同时，银表面上可以测到很强的苯硫酚的SERS信号。为了减少抗体分子与基底银间的这种非特性吸附作用，我们尝试以小牛血清蛋白BSA封闭基底的活性位，取得了很好的效果，使银表面的SERS强度明显减弱。更多还原

【Abstract】 Surface-Enhanced Raman Spectroscopy(SERS)has been extensively applied into various microanalysis on molecular level because of its high sensitivity and it will have great practical potential in Labeled Immunoassay especially when NIR FT-Raman Spectrometer is used as detector. In this article SERS labeled immunogold is made out for immunoassay:The relationship between FT-SERS intensity and the size of spheroid gold nanoparticles is firstly discussed with thiophenol as probe molecule. Perfect SERS signal was obtained from NIR FT-Raman Spectrometer when 50nm Au nanoparticles were prepared by Hydroxylamine Seeding, while no SERS can be obtained on 30nm Au nanoparticles. It is also found that the size growth of self-assembled Au nanoparticles can also increase SERS intensity.The above 50nm Au sol was then made into SERS Reporter-thiophenol labeled immunogold. The amount of the reporter molecule, the reaction time between thiophenol and gold and the combination of thiophenol labeled gold with goat anti-rat IgG are discussed.SERS labelled immunogold was then used as probe to detect the interaction between the Goat anti-rat IgG and silver substrate (which we will use it as the substrate for solid antibody). It is found from experiment that SERS labelled immunogold can be non-specifically adsorbed onto silver surface uniformly because of the strong interaction between the protein molecule and silver. Strong SERS signal of thiophenol was also observed after the silver was treated with thiophenol labelled immunogold. BSA was used to eliminate the non-specific adsorption.更多还原