The Preliminary Studies On Separation Of Proteins By Capillary Electrophoresis

The Capillary Electrophoresis is a useful technology in separation and analysis of bio-materials, which have get more and more attention for its high performance. It developed rapidly after the end of 1980s and was used in varied fields such as pharmaceutical analysis, separation of chirality's matter and bio-macromolecule. Our laboratory bought a HP3D Capillary Electrophoresis apparatus recently in order to analyze bio-macromolecule from plant. There is little information about analysis of plant proteins by this way in domestic. So the purpose of this article is to master principle and instrument operation and to set up the way to analyze water-soluble proteins. This is the base of further research.First, the appropriate condition of Capillary Electrophoresis was found after separation of protein-marker and bovine serum albumin. The effect of capillary and temperature was tested. The result showed that the capillary with 50cm length and inter diameter 50 μn was appropriate and the best temperature was from 25℃ to 35℃. The relations between injection and ultraviolet absorption were analyzed at the same time.Then the separation method of water-soluble proteins through capillary zone electrophoresis (CZE) was studied. The effect of pH of phosphate buffer on separation of water-soluble proteins was tested. The results indicated that the pH2.5 buffer was the suitable one. The best measure wavelength was found in the 3-D chromatogram.The separation of water-soluble proteins from egg and Pinus tabulaeformis Carr. ovule was done by capillary zone electrophoresis (CZE) on the base of success in separation of single protein sample (bovine serum albumin), and the electrophoresis condition was set finally.And the separation of water-soluble proteins was done with micelle electro kinetic capillary chromatography (MECC). The protein adsorption increased and the efficiency of separation declined when the SDS was added in phosphate buffer.Additionally, the effect of some factors which have tight relation with electrophoresis is discussed.