For New Methods Of Preparation Of Sugar Chip

BackgroundFor the last decade, we recognized that the interaction between the glycans on the surface of cells and the proteins promoted the sticky between cell and cell, and participate the development of vertebrate. This kind of protein which interact with glycan called glycan-binding protein, GBP. Specific protein-glycan interactions underlie many aspects of important biological processes, for example, cell differentiation, cell adhesion, immune response, trafficking and tumor cell metastasis, which occurs through glycans of glycoprotein and glycolipid, as well as polysaccharide displayed on cell surfacedefectglycan binding proteins (GBPs). Glycan-protein interactions play important biological roles in biological processes. But there is a lack of simple high-throughput methods to elucidate recognition events between carbohydrates and protein. Especially defect a study tool similar to the DNA microarray or protein microarray. Nowadays, the study of glycan microarray is just on the first stage. The establishment of the platform of glycan microarray will provide a high-throughput methods for the study of the relations between the glycans and proteins.AimStudy the preparation of the glycan microarray.MethodsThis paper reported a simple, convenient and efficient method for preparing glycan microarrays based on hydroxyl-modified surfaces on glass support. Here, two strategies have been developed for the production of glycan microarrays by the underivatized oligosaccharides were efficiently immobilized on hydroxyl-functionalized glass surface by formation of glycosidic bonding with the hemiacetal group at the reducing end of the suitable carbohydrates via condensation. Firstly, untreated glass slides were amino- and epoxy-silanized, respectively. Then, they were further hydroxyl activated with different surface chemistries.Results Overall best results in terms of conveniency and sensitivity were obtained with hydroxyl- activation on epoxysilanized surfaces.Develop a simple, convenient and efficient method for preparing glycan microarrays based on hydroxyl-modified surfaces on glass support. In this thesis, spotting concentration, carbohydrates immobilization substrates, target labeling and affinity reaction condition were investigated, and the carbohydrates chip of liver-protein researches were applied. The major contributions include the following aspects:1. Overall best results in terms of conveniency and sensitivity were obtained with hydroxyl- activation on epoxysilanized surfaces.a. The result showed the fluorescent intensity was the highest when spotting buffer was acetate buffer.b. 3% TopBlock solution was used as blocking reagent in all experiments reported here.c. The result showed the fluorescent intensities were higher under neutral and alkalescent condition.d. The best incubation time is 1h. The best incubation temperature is 4℃.e. The best spotting concentration is 1mmol/mL. f.The best storage condition is 4℃.2. Study the glycan-binding protein of the liver with the technology of glycan microarray. Recognized that the difference of the distribution and quantity of the liver glycan-binding protein at the different developmental stage and different tissue, settle the foundation for the study of liver glycan-binding protein.3.Study the antiglycan antibody of the health human serum with the technology of glycan microarray. Settle the foundation for the study of antiglycan antibody of the health human serum.