摘要
近几年来共振光散射技术逐渐被应用于生物分子检测中,以及纳米金在共振光散射分析技术中应用的研究也越来越受到关注。
本文主要研究了IgG免疫复合物微粒的共振散射光谱。结果表明,在PBS缓冲溶液中,免疫球蛋白G(IgG)与羊抗人免疫球蛋白G发生特异性结合,形成了抗原-抗体免疫复合物微粒,使相应的各同步散射峰的共振散射强度大大增强。据此建立了检测免疫球蛋白G的方法,此方法操作简便、快速。
进一步研究了金标记免疫共振散射光谱法检测人免疫球蛋白IgG的方法。研究结果表明,这种新型的免疫分析方法灵敏度高,检出限低,检测快速。
Resonance light scattering (RLS) technique is a new molecular spectrum analytical method. With the distinct advantages of convenience , sensitivity and selectivity, RLS is increasingly gaining more and more attention among the international and internal analytical and biochemical researchers. During the ten plus years, especially with the development of the molecular recognition functional RLS probes, the high-sensitivity RLS technique is developed even more quickly and play an important role in the biochemical analysis process, demonstrating enormous application potential To further study and develop high-sensitivity, high-selectivity, simple and convenient RLS new determination method and system naturally becomes a very important study subject.
This thesis contains three sections. Section one is a review, which involves the history, the basic principles, characteristics and the main systems of RLS, and applications of RLS systems in pharmaceutical analysis, and principles and applications of immunoassay. We also summarized the recent developments of gold nanoparticles in RLS .
In the second section, it has been found that the resonance scattering spectral study of IgG immunocoplex particles and its analytical application. In pH 7.4 PBS buffer solution, fibrinogen would combine with its antibody and produce immunocomplex particles that resulted in resonance scattering (RS) effect at 310 nm. The intensitiesΔI_(RS) at 310 nm are proportional to the fibrinogen concentrations from 1.8μg·mL~(-1) to 100μg·mL~(-1) ; The detection limits is 0.92μg·mL~(-1). This method has the advantages of simple operation, high sensitivity and fast response. The method has been applied to the determination of human IgG, with satisfactory results.
In the third section, we developed a novel and sensitive analysis method that nanogold-labeled immunoresonance scattering spectral assay for trace IgG. Nanogold in size of 13nm was prepared by trisodium citrate method and was labeled with goat anti-human IgG. The immune reaction between nanogold-labeled antibodies and antigens took place in pH 7.4 PBS buffer solution that resulted in the aggregation of the nanogold,and the intensity of resonance scattering peaks at 580 nm enhanced greatly. The enhanced intensityΔI_(RS) is proportional to the concentrations in the range of 0.45-300μg·mL~(-1) for IgG. The detection limits are 0.37 ug/ml (3σ) . This method has been successfully applied to the determination of HIgG and showed extensive potential applications in analytical and bioanalytical chemistry.
引文
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