Physico-chemical characterization of proteins by capillary electrophoresis.
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-Lactoglobulin exhibited increasingly lower mobilities than those expected of boric acid/borate at pH 7.0 and 8.0 (I=20 mmol l(-1)).
Rabiller-Baudry
M
M
Laboratoire des Procédés de Séparation, UA Université de Rennes 1-INRA, France.
Bouguen
A
A
Lucas
D
D
Chaufer
B
B
D001888:Boric Acids; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004563:Electrochemistry; D004573:Electrolytes; D019075:Electrophoresis, Capillary; D006863:Hydrogen-Ion Concentration; D007768:Lactalbumin / Q000737:chemistry; D007782:Lactoglobulins / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D009994:Osmolar Concentration; D010710:Phosphates; D017680:Potassium Compounds; D011506:Proteins / Q000737:chemistry*
D001888:Boric Acids; D004573:Electrolytes; D007782:Lactoglobulins; D010710:Phosphates; D017680:Potassium Compounds; D011506:Proteins; D007768:Lactalbumin; C013216:potassium phosphate; D009113:Muramidase; C032688:boric acid; C018279:sodium phosphate