Micropreparative solution isoelectric focusing of peptides and proteins in nonwoven strip

0386015 - ÚIACH 2013 RIV CZ eng C - Conference Paper (international conference)
Duša, Filip - Šlais, Karel
Micropreparative solution isoelectric focusing of peptides and proteins in nonwoven strip.
CECE 2012. 9th International Interdisciplinary Meeting on Bioanalysis. Brno: Ústav analytické chemie AV ČR, v. v. i, 2012 - (Foret, F.; Křenková, J.; Guttman, A.; Klepárník, K.; Boček, P.), s. 66-68. ISBN 978-80-904959-1-3.
[CECE 2012. International Interdisciplinary Meeting on Bioanalysis /9./. Brno (CZ), 01.11.2012-02.11.2012]
R&D Projects: GA MV VG20102015023; GA MŠMT(CZ) EE2.3.20.0182
Institutional support: RVO:68081715
Keywords : isoelectric focusing * preparative * whey
Subject RIV: CB - Analytical Chemistry, Separation

Recently we devised a new instrument for micropreparative analysis [1]. It is based on solution phase isoelectric focusing (sIEF) performed in a narrow channel (approximately 2 – 4 mm wide) with a strip of nonwoven fabric serving as the analysis bed. The strip can be precut before an analysis to make harvesting of fractions easier. Isoelectric focusing is driven by a programmable electrophoretic power supply. Progress of analysis is monitored by addition of colored isoelectric point (pI) markers. Usually sIEF runs are performed overnight. Evaporation of water is one of crucial features of analysis and it forces free liquid to shrink into the nonwoven fabric strip. Moreover, water evaporation increases liquid viscosity by altering ethylene glycol/water ratio. Fractions can be harvested simply by collecting and washing precut segments after the sIEF run. In this paper we used sIEF device for purification of caseinomacropeptide (CMP) from crude whey. Obtained fractions were further analyzed by HPLC. From acquired data a plot with chromatograms of all fractions was constructed to show purification profile of CMP. We proved that the new instrument is capable of quantitative purification of CMP complex from a globulin fraction.
Permanent Link: http://hdl.handle.net/11104/0215261