通用稀土纳米颗粒标记的构建方案及其应用研究

Lanthanide labeling bioassay has a high throughput, but the method is not sensitive enough for the low-abundant proteins quantification and early detection of cancers. A viable option to improve the analytical sensitivity is to increase the number of elemental tags using a universal labeling strategy. The present proposal aims at (1) developing a universal lanthanide-nanoparticle-based labeling method for bioassays: to synthesize or select a series of nano-cage structures as synthetic templates and carriers for the lanthanide nanoparticles, and to simplify the synthesis and labeling procedures to facilitate the radioactive labeling; (2) conducting lanthanide-nanoparticle-based high-throughput protein quantification and immunoimaging with an integrated analytical approach using the mega-science facilities, analysis platforms, and imaging equipments in our institute; (3) investigating the effects of the physicochemical properties of nanoparticle-tags on their metabolic behaviors and toxicological consequences in vivo, in order to evaluate the practicality and safety of lanthanide-nanoparticle-based labeling method for in vivo imaging and disease diagnosis/therapy. This project, depending on distinguishing features of a cross-discipline of analytical science and nanoscience, will boost the applications of nuclear analytical techniques in protein analysis, immunoimaging and disease diagnosis/therapy.

基于稀土元素标记的分析方法有很好的分析通量，但方法的灵敏度不能满足低丰度蛋白质定量及肿瘤早期诊断的需求。在保证标记方法适用于多数稀土元素的前提下提高标记原子的个数，是一个提高分析灵敏度的可行途径。本项目旨在（1）建立一套通用的稀土纳米颗粒标记方法：合成或选用一系列纳米笼形结构作为稀土纳米颗粒的合成模板和载体，简化合成与标记环节,便于实现放射性标记；（2）综合运用本单位内多个大科学装置、分析平台、成像设备，开展稀土纳米颗粒标记技术在高通量蛋白质定量、免疫分析成像等领域的应用研究；（3）研究标记单元的物化性质对其体内代谢行为及纳米毒理效应的影响，评估稀土纳米颗粒标记技术应用于活体成像、疾病诊疗等领域的可行性与安全性。本项目将立足于分析科学与纳米科学的学科交叉，推动核分析技术在蛋白分析、免疫成像、疾病诊疗等领域的应用。

经过4年的实施，我们已经建立了通用的稀土纳米颗粒标记方法，利用同样的笼形结构模板，可以方便的合成多种纳米稀土颗粒标记体。利用稀土纳米颗粒标记方法建立的蛋白免疫分析，其检出限比稀土离子标记降低了1个数量级。我们利用XRF和LA-ICP-MS技术对组织中或固相免疫吸附表面上的纳米颗粒标记体进行分布成像或定量研究。我们的工作表明，利用纳米稀土颗粒进行免疫标记，并配合ICP-MS、LA-ICP-MS、XRF和NanoSIMS检测技术，可以构建一种高灵敏、高通量的蛋白分析方法。项目发表论文17篇，专著章节3部。