Informace o publikaci

Micro fractionation of proteins and peptides by solution phase isoelectric focusing in nonwoven fabric strip


DUŠA Filip ŠLAIS Karel

Rok publikování 2012
Druh Konferenční abstrakty
Popis Here we suggest a new device for solution phase isoelectric focusing of sub milliliter sample volumes. Device is set up from series of segments of nonwoven fabric aligned into a strip in an open channel located in a cool box. Isoelectric focusing is driven by programmable electrophoretic power supply. Crucial feature of the new method is the evaporation of water during isoelectric focusing run which shrinks all free liquid into the strip and moreover it increases the liquid viscosity due to the change in ethylenglykol/water ratio in the separation medium. Final pH in strip segments is monitored by the position of colored pI markers synthetized in our laboratory. Fractions are harvested by simple collecting and washing of segments of the strip. Obtained liquid fractions are further analyzed by micro reverse phase liquid chromatography. This new method is applied for analysis of crude whey peptides and proteins. It was proved that the new simple method quantitatively separates the caseinomacropeptide complex from the globulin fraction.