Mise au point d'une technique de séparation analytique des protéines : électrophorèse multiphasique en présence d'espaceurs hautement diversifiés
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Abstract EN:
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Abstract FR:
In multiphasic electrophoresis, dispersions due to diffusion and electrophoretic migration are counteracted by moving boundaries. The introduction of highly diversified spacers can generate a moving electric field strength gradient, in which proteins can be separated according to their electrophoretic mobility. Our investigations were led according to two main directions: i) isotachophoresis with highly diversified spacers, in a support media generating an electroosmotic counterflow; ii) utilization of snow and telescope electrophoretic systems, with highly diversified spacers. The experimental study of the influence of the important parameters (particularly the nature and the concentration of ionic species in each phase) was in good agreement with the theory. Resolution power was evaluated by revelation of particular proteins and by measurements of local electric field strength an pH at the end of the experiments. This work leads to the development of a technique of protein separation, which is very easy and fast to set up and has a resolution power quite comparable to that of isoelectric focusing. Further, it allows protein analysis at pH differing from their isoelectric point. Finally, we show the new axes of investigation which have to be developed to improve the resolution power