Non Polar Electrical Field Flow Fractionation (Np-EFFF)
The purpose of the Non Polar Electrical Field Flow Fractionation (Np-EFFF) project is to explore a novel method of isolating membrane and lipophilic proteins from tissue samples so that they can be more fully characterized. Presently existing methods rely on waterbased solvents and the use of surfactants to suspend the proteins in solution. This process can destroy the protein function. The nonpolar solutions are expected to maintain the native conformation and function of the proteins and to permit the study of their function. We will design and construct the NpEFFF instrument which comprises two oppositely charged plate electrodes, a thin ribbonlike channel defined between the plates, a means for delivering a flowing medium through the channel, and a protein detector at the end of the channel. Protein fraction is extracted from a tissue sample and the extracted material is suspended in the non-polar solvent. In the EFFF channel, proteins are attracted to the plate of opposite electrical charge and the progress through the channel depends on the size and charge of the proteins. If successful, this method will allow for the understudied proteins that are critical to many life functions to be isolated and characterized.