Capillary electrophoresis of seed 2S albumins from Lupinus species

Journal of Chromatography. a
B P Salmanowicz

Abstract

Two modes of capillary electrophoresis (CE)--free-solution capillary zone electrophoresis (CZE) and sodium dodecyl sulfate capillary electrophoresis (SDS-CE) using a non-gel sieving matrix--have been developed for comparative analysis of low-molecular-mass 2S albumin isoforms from lupins. The albumin fraction and 2S albumins were separated in uncoated fused-silica capillary by CZE with 0.02 M phosphate buffer, pH 7.3, containing the sodium salt of phytic acid. The use of phytic acid (0.025 M) as buffer modifier and ion-pairing agent improved migration reproducibility, peak shape and separation efficiency. The reduced 2S albumins were separated by SDS-CE using a high concentration (0.3-0.5 M) mixture of tris(hydroxymethyl)aminomethane and borate buffers in uncoated fused-silica capillary. Of the various polymers used as non-gel sieving matrix, SDS-CE with a 10% dextran solution was found to be suitable for separation of 2S albumin polypeptides with molecular masses of 4,000-7,000 and 8,000-11,000. The addition of glycerol or ethylene glycol to the SDS separating buffer improved the resolution of polypeptides. The examined Lupinus species showed species-specific CZE and SDS-CE migration profiles of the 2S albumins.

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Citations

Feb 5, 2002·Electrophoresis·V Dolnik, K M Hutterer
Nov 16, 2001·Phytochemical Analysis : PCA
Jun 19, 2012·Analytical and Bioanalytical Chemistry·Angela R Piergiovanni
Feb 11, 2005·Electrophoresis·Julia A FruetelDebbie Padgen
Jul 2, 2002·Analytical Chemistry·Shen Hu, Norman J Dovichi
Aug 18, 2005·Journal of Agricultural and Food Chemistry·Angela R Piergiovanni, Giovanni Taranto

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