基于纳米标记与传感阵列的磷酸化蛋白高灵敏高通量检测新方法

Protein phosphorylation is a major regulatory mechanism that controls almost all life activities of each basic cellular process. Accordingly, phosphorylation analysis, in particular the quantitative measurement of phosphoproteins becomes a hot topic of many research areas including Life Science, Chemistry and Medicine. In light of some difficulty issues of current methods for phosphorylation analyses such as complex operation, expensive cost, sensitivity and accuracy is required to be improved, this project will establish nanoparticles-based labels and nano-biosensor arrays for measurement of phosphoproteins and related biomarkers with high sensitivity and high throughput. This research will not only explore the dynamic mechanism of molecular recognition between the probe and the phospho-biomarkers, but also discuss the adsorption and electronic transfer mechanism of proteins in nano-interface. The study in this proposal also includes quantitative analysis of the sites of phosphoproteins and new technology for simultaneous detection of multiple phospho-biomarkers. Combined with flow injections, test strip and screen printing technology, this project is primary aimed at developing sensitive, low-cost, simple and practical diagnostic tools for phospho-biomarkers and will potentially produce better immunoassay method and novel devices. We believe that the implementation of this project will establish new technology for quantification of phosphoproteins and phospho-biomarkers, which has great clinical significance in early diagnosis and treatment of cancer and other disease.

蛋白质磷酸化几乎调节着生命活动的每一过程，围绕着磷酸化蛋白的研究已成为生命科学、化学、医学等领域一个引人注目的焦点。本项目针对目前磷酸化蛋白检测方法操作复杂、价格昂贵、灵敏度与准确性尚需提高等难点，将构筑纳米生物标记物与纳米生物传感阵列，以实现基于纳米界面体系的磷酸化蛋白及关键生物标志物的高灵敏高通量检测。探讨探针分子与磷化蛋白及相关生物标志物的结合动力学过程，阐明蛋白质分子在纳米界面的吸附、识别与电子传递等作用机制。开展磷酸化蛋白质的多磷化位点定量分析与多态性分析的研究，建立多种磷化蛋白相关生物标志物的联合检测新技术。结合流动注射、免疫试纸条与丝网印刷电极技术开发高灵敏、低成本、简单实用的快速测定系统，以期获得生物标志物免疫分析新方法与新器件。相信通过本项目的实施，将建立磷酸化蛋白及关键生物标志物的定量测定新技术，对于癌症和重大疾病的早期诊断和治疗具有非常重要的科学意义和临床价值。