蛋白质是生命科学中一类重要的生物大分子，是生物体实现其功能的物质基础，因此蛋白质的高灵敏度检测和分析一直是研究热点。针对毛细管电泳检测灵敏度因检测光程短而较低的问题，拟构建新型基于温度控制蛋白质捕捉和释放的在线固相萃取-毛细管电泳体系，用于蛋白质的富集和分析。首先在柱上固相萃取硅胶整体柱上键合单层聚N-异丙基丙烯酰胺（PNIPAM），利用该聚合物在高于临界溶解温度呈疏水性和在低于临界溶解温度呈亲水性的特性进行蛋白质的捕捉和释放。通过优化在线固相萃取柱长度、硅胶整体柱孔隙率以及提高PNIPAM键合密度，并将固相萃取和场放大效应结合，实现在柱上固相萃取-毛细管电泳体系上蛋白质的高效富集和分离。此外我们还将构建上述在线固相萃取-毛细管电色谱体系，将固相萃取、场放大效应和色谱区带压缩效应相结合，提高蛋白质的检测灵敏度。通过上述的研究，我们旨在发展新型高效蛋白质浓缩和分离方法。

Proteins are important biomacromolecules in life science. They are material basis for organisms to fulfill their functions. Therefore, high-sensitivity analysis and detection of proteins have attracted great attention for decades. In order to improve relatively low detection sensitivity of capillary electrophoresis due to small sample volume and short optical length for on-line detection, we plan to design a novel temperature responsive capture and release of proteins on-capillary solid phase extraction-capillary electrophoresis for enrichment and analysis of proteins. A monolayer of poly(N-isopropylacrylamide) (PNIPAM) is to be immobilized on silica monolithic column. This polymer exhibits hydrophilicity below critical solution temperature and hydrophobicity above critical solution temperature. Accordingly, proteins could be captured on the monolayer of PNIPAM above the critical solution temperature and could be released below the critical solution temperature. By optimizing solid phase extraction column length, porosity of silica monolithic column and increasing PNIPAM bonding density as well as combining solid phase extraction and field amplified sample stacking technique, detection sensitivity of proteins could be greatly improved on this system. In addition, we will construct aforementioned solid phase extraction column-capillary electrochromatography for efficient enrichment and analysis of proteins. Solid phase extraction, chromatographic zone-sharpening effect and field amplified sample stacking are to be utilized to improve protein detection sensitivity. Based on the above research, we aim to develop novel and efficient methods for protein concentration and separation.