Preliminary Study Of Proteomic Separation Technologies Based On Capillary Electrophoresis

It is the object to develop complex proteomic separation technologies based ontwo-dimensional capillary electrophoresis in this dissertation. Specially designedhollow fiber interfaces are used to construct separation platforms of 2D CIEF-CGEand 2D CIEF-CZE. Hemoglobin variants are studied by 2D CIEF-CGE, as well asribonuclease, by 2D CIEF-CZE with dynamic focusing and reversed electroosmoticflows. Unique abilities of concentration and highly separation efficiencies areexhibited in these novel systems.After the investigation on the electrophoresis of proteins with extreme pIs in apH gradient, a new mode of CE, "pH gradient driving electrophoresis"(PDGE), isset up applying the technique of previously established pH gradient plus partlyinjection. Possible protocols to improve the separation methods for proteins withextreme pIs are given by theoretical study on the mass transferring of PDGE andexperiment of separating egg white solution.To overcome the disadvantage origined from the addition of carrier ampholytesin CIEF, a monolithic immobilized pH gradient (M-IPG) is designed and prepared tosupply possible research methods improving resolution and matchability of detectors.M-IPG is utilized in CIEF to give results similar to that in free solution and indicategood resolution and efficiencies simultaneously. Perfect results are obtained whileusing this column to separate real samples of antler and rat liver extract.