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Original title:
Micropreparative solution isoelectric focusing of peptides and proteins in nonwoven strip
Authors: Duša, Filip ; Šlais, Karel
Document type: Papers
Conference/Event: CECE 2012. International Interdisciplinary Meeting on Bioanalysis /9./, Brno (CZ), 2012-11-01 / 2012-11-02
Year: 2012
Language: eng
Abstract: 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.
Keywords: isoelectric focusing; preparative; whey
Project no.: VG20102015023 (CEP), EE2.3.20.0182
Funding provider: GA MV, GA MŠk
Host item entry: CECE 2012. 9th International Interdisciplinary Meeting on Bioanalysis, ISBN 978-80-904959-1-3
Note: Související webová stránka: http://hdl.handle.net/11104/0215261
Institution: Institute of Analytical Chemistry AS ČR (web)
Document availability information: Fulltext is available in the digital repository of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0215261
Permalink: http://www.nusl.cz/ntk/nusl-135528
The record appears in these collections:
Research > Institutes ASCR > Institute of Analytical Chemistry
Conference materials > Papers
Authors: Duša, Filip ; Šlais, Karel
Document type: Papers
Conference/Event: CECE 2012. International Interdisciplinary Meeting on Bioanalysis /9./, Brno (CZ), 2012-11-01 / 2012-11-02
Year: 2012
Language: eng
Abstract: 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.
Keywords: isoelectric focusing; preparative; whey
Project no.: VG20102015023 (CEP), EE2.3.20.0182
Funding provider: GA MV, GA MŠk
Host item entry: CECE 2012. 9th International Interdisciplinary Meeting on Bioanalysis, ISBN 978-80-904959-1-3
Note: Související webová stránka: http://hdl.handle.net/11104/0215261
Institution: Institute of Analytical Chemistry AS ČR (web)
Document availability information: Fulltext is available in the digital repository of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0215261
Permalink: http://www.nusl.cz/ntk/nusl-135528
The record appears in these collections:
Research > Institutes ASCR > Institute of Analytical Chemistry
Conference materials > Papers
Record created 2013-01-04, last modified 2023-12-06