Investigation Of Interactions Between Heparin-like Sulfated Polysaccharides And Proteins At Both Molecular And Cellular Level

Sulfated polysaccharides play a central role in many biological processes including signaling, cell growth and differentiation through interaction with proteins. Investigation of the interactions is important in understanding biochemical and physiological processes, and can also accelerate the development of new carbohydrate-based drugs. This thesis studied the interactions between heparin-like sulfated polysaccharides and proteins including granulocyte colony-stimulating factor (G-CSF) by capillary electrophoresis (CE), high performance liquid chromatography (HPLC), mass spectrometry (MS) and flow cytometry (FC), etc.The interactions between heparin and different types of proteins were investigated by capillary zone electrophoresis (CZE). It was found that CZE could be used in both slow and fast on-and-off kinetic systems. The binding constants of granulocyte-macrophage colony-stimulating factor (GM-CSF) with heparin were determined for the first time. It also found that GM-CSF could specifically interact with heparin.We systematically investigated the interactions between G-CSF and heparins with different chain length, different source and different sulfate position by CZE both qualitatively and quantitatively. It was found for the first time that G-CSF could bind to heparin and the binding was dependent on the chain length, the sulfate position and the source of heparin. A CZE method was developed for the separation of a heparinase degraded oligosaccharides compatible to study the interactions of the heparin oligosaccharides with G-CSF. Heparin oligosaccharides could be baseline separated quickly without any additives at normal polarity by...