Cyclodextrin aided separation of peptides and proteins by capillary zone electrophoresis.
Carboxymethylated-beta-cyclodextrin (CMBCD) in the electrophoretic medium (aqueous 50 mM sodium phosphate, pH 2.5) enhanced the separation using raw fused-silica capillaries in CZE of the four standard proteins: alpha-chymotrypsinogen A, cytochrome c, lysozyme and ribonuclease A. Furthermore, with 20 mM CMBCD in the electrophoretic medium, the cis-trans isomers of angiotensin could be separated at room temperature, whereas the separation of the conformers required subambient temperatures as low as -20 degrees C without CMBCD in the electrophoretic medium [50 mM sodium phosphate (pH 2.5), containing 10% (v/v) methanol]. Addition of heptakis(2,6-di-O-methyl)-beta-cyclodextrin (DMBCD) had no effect on the separation of the above proteins and peptides. The results suggest that in microcolumn separation techniques, certain cyclodextrin additives can be useful selectivity enhancers.
Rathore
A S
AS
Department of Chemical Engineering, Yale University, New Haven, CT 06520-8286, USA.
Horváth
C
C
D000595:Amino Acid Sequence; D000803:Angiotensin I / Q000302:isolation & purification*; D002919:Chymotrypsinogen / Q000302:isolation & purification; D003080:Cold Temperature; D003505:Cyclodextrins / Q000737:chemistry*; D003574:Cytochrome c Group / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D009113:Muramidase / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D012259:Ribonuclease, Pancreatic / Q000302:isolation & purification; D047392:beta-Cyclodextrins
D003505:Cyclodextrins; D003574:Cytochrome c Group; D011506:Proteins; D047392:beta-Cyclodextrins; C038119:heptakis(2,6-O-dimethyl)beta-cyclodextrin; D002919:Chymotrypsinogen; D000803:Angiotensin I; D012259:Ribonuclease, Pancreatic; D009113:Muramidase; C031215:betadex