Physico-chemical characterization of proteins by capillary electrophoresis

Journal of Chromatography. B, Biomedical Sciences and Applications
Murielle Rabiller-BaudryBernard Chaufer

Abstract

The electrophoretic mobility of proteins was successfully determined by means of capillary electrophoresis (CE) with various background electrolytes (BGEs). The objective was focused on the variation in BGE physico-chemical composition and the consequential impact on the observed protein charge. Experimental and calculated mobilities, according to Henry's equation, versus ionic strength have been compared. For positively-charged lysozyme, a good agreement between observed and calculated mobilities was observed using triethanolamine chloride at pH 7.0 as the BGE. Mobility close to zero was shown using borate (pH 8.0) and phosphate (pH 7.0) at a low ionic strength of about 20 mmol l(-1), and as a consequence, specific adsorption of oxyanions was evidenced. Lysozyme retention in the case of reversed-phase high-performance liquid chromatography (RP-HPLC) was decreased by the presence of phosphate ions. CE and HPLC are complementary tools for characterizing the behaviour of lysozyme. On the other hand, the mobility of the negatively-charged alpha-lactalbumin remained constant as regards phosphate at pH 7.0 in the 20-200 mmol l(-1) range, contrary to the decrease that had been expected with the increasing ionic strength. beta-Lactogl...Continue Reading

References

Sep 7, 1990·Journal of Chromatography·J K Towns, F E Regnier
Dec 6, 1994·Proceedings of the National Academy of Sciences of the United States of America·J GaoG M Whitesides
Oct 15, 1993·Journal of Chromatography. a·N de JongC Olieman
Feb 20, 1990·Biotechnology and Bioengineering·M T Tyn, T W Gusek

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Citations

Nov 25, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Murielle Rabiller-Baudry, Bernard Chaufer
Apr 10, 2007·Analytical Biochemistry·Earle StellwagenNancy C Stellwagen
Jun 26, 2012·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Anang SedyohutomoChuzo Fujimoto
Apr 23, 2013·Journal of Food Science·J C BanachB P Lamsal
Dec 22, 1999·Electrophoresis·V Kasicka
May 15, 2003·Journal of Agricultural and Food Chemistry·Jose E N Gutierrez, Lisa Jakobovits

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