临床上的癌症标志物多是糖蛋白，因为癌症与蛋白质糖基化密切相关。由于糖蛋白存在诸多亚型和复杂糖修饰，给分离分析带来巨大挑战，迫切需要发展新技术对其开展精准研究。本项目选择血液和唾液作为研究对象，通过发展新型高分辨双向差异凝胶电泳技术，对凝集素富集的糖蛋白进行分离分析，建立糖蛋白标志物发现新技术；选择目标糖蛋白进行抗体免疫富集，结合双向凝胶电泳建立糖蛋白亚型和糖链结构分离鉴定新技术；将发展的新技术用于发现胃癌和肺癌的糖蛋白标志物。本研究意义在于：第一，新型双向凝胶电泳技术将促进蛋白质组学研究；第二，新技术将弥补常规方法对糖蛋白亚型难以区分的缺点；第三，糖蛋白糖链结构分离鉴定新技术有助于实现标志物的精准定性；第四，新技术的应用将推动癌症分子诊断的发展。通过项目研究，有望解决糖蛋白标志物分离鉴定困难的现状，提高其诊断癌症的特异性和灵敏度，为糖蛋白标志物的发展提供技术支持。

Many clinically used cancer biomarkers are glycoproteins, due to the fact that glycosylation is closely related to carcinogenesis. Glycoproteins usually have many isoforms along with complex glycans, bringing great challenges to bioseparation and analysis. Therefore, it is urgent to develop new technologies for the accurate identification of glycoprotein biomarkers. In this project, blood and saliva samples are selected as targets for research. Through developing novel and high resolution two-dimensional difference gel electrophoresis, glycoproteins enriched by lectins will be separated and analyzed. Thus new technology will be established for the discovery of glycoprotein biomarkers. Antibody immunoprecipitation is employed to enrich target glycoproteins. Different isoforms of target glycoprotein with various glycans will be separated and characterized. Combining the developed new technologies, the project will find glycoprotein biomarkers for gastric cancer and lung cancer. The values of this project are as follows. Firstly, novel two-dimensional gel electrophoresis will benefit proteomics studies. Secondly, new technology will overcome the shortcoming of routine method that cannot separate the isoforms of glycoproteins. Thirdly, the newly developed technology of glycan structure characterization could lead to the precise identification of biomarker. Fourthly, newly developed technologies will contribute to the development of cancer molecular diagnostics. Through the study of this project, it is expected to solve the key issues in the separation and identification of glycoprotein biomarkers, which will ultimately improve the specificity and sensitivity of cancer detection. The achievement of this project will provide key technology support to the discovery of novel glycoprotein biomarkers.