葡萄糖转运体1（GLUT1）是12次跨膜疏水性蛋白质，难以分离纯化并保留活性构象，转运机制复杂；细胞膜表达靶蛋白数量有限；固定化细胞毛细管柱制作技术不易掌握、使用寿命短、与MS联用难等问题，使以膜蛋白为靶点的固定化细胞毛细管电泳（ICCE）筛选方法受到挑战。本项目旨在构建人GLUT1真核表达质粒，建立高表达GLUT1的HEK293细胞，在生理或接近生理条件下，以DMSO为对照样品，进行一系列新技术探索，定性定量研究葡萄糖等阳性物质与活HEK293细胞、固定HEK293细胞、超表达GLUT1的活HEK293细胞、超表达GLUT1的固定HEK293细胞的相互作用，并计算结合动力学参数，建立以GLUT1为靶点NICCE筛选药物新方法。筛选合成糖类衍生物和天然产物，进行细胞增殖和细胞毒性实验。本研究将有效解决上述问题，为体内抗肿瘤药效实验和发现源头创新药物奠定基础、建立新筛选平台并提供筛选新思路。

The human glucose transporter GLUT1 is an twelve-transmembrane and extremely hydrophobic protein. However, as to extremely hydrophobic proteins, there exists a serious problem, i.e., isolating and purifying these proteins from the cell membrane will typically results in loss of native conformation, and it’s transport mechanism is complicated. Additionally, a number of target protains expressed on cell membranes is limited; the preparation technology of the immobilized cell capillary column is not easy to be mastered and it has a relatively short service life, also, it is difficult to couple the immobilized cell capillary electrophoresis (ICCE) with mass spectrometry. These factors make the method of screening ligands with the cell membrance receptors as targets by ICCE meet new challenges. In the present study, a novel method is proposed for the first time, in which, firstly, the GLUT1-eGFP plasmids will be transferred into HEK293 cells using electrotransfection. Secondly, under the physiological condition or approximately physiological conditions, a series of new technologies about this method will be investigated using DMSO as control sample. Finally, the novel non-immobilized cell capillary electrophoresis (NICCE) will be investigated and established by optimizing a series of parameters of the HPCE, and the interactions of GLUT1 with glucose and some other positive compounds will be studied by qualitative-quantitative analysis metnod and the defferent cells (living HEK293 cells, paraformaldehyde-fixed HEK293 cells, overexpression GLUT1 living HEK293 cells and overexpression GLUT1 paraformaldehyde-fixed HEK293 cells). The kinetic parameters of interaction between GLUT1 and glucose/other positive compounds will be calculated by the model. And the new NICCE screening platform will be established and used to rapidly screen a series of the new synthetic carbohydrate derivatives and the natural products. Furthermore, the efficacy tests of cell growth and cytotoxicity will be carried out for these binding-active compounds. This study will solve the above-mentioned problems effectively, lay the foundation for discovering the source of innovative drugs and the efficacy tests of anti-tumor in vivo, and establish a new method and new technology for screening anti-tumor drugs.