On-line hyphenation of capillary isoelectric focusing and capillary gel electrophoresis by a dialysis interface.
An on-line two-dimensional (2D) capillary electrophoresis (CE) system consisting of capillary isoelectric focusing (CIEF) and capillary gel electrophoresis (CGE) was introduced. To validate this 2D system, a dialysis interface was developed by mounting a hollow fiber on a methacrylate resin plate to hyphenate the two CE modes. The two dimensions of capillary shared a cathode fixated into a reservoir in the methacrylate plate; thus, with three electrodes and only one high-voltage source, a 2D CE framework was successfully established. A practical 2D CIEF-CGE experiment was carried out to deal with a target protein, hemoglobin (Hb). After the Hb variants with different isoelectric points (pIs) were focused in various bands in the first-dimension capillary, they were chemically mobilized one after another and fed to the second-dimension capillary for further separation in polyacrylamide gel. During this procedure, a single CIEF band was separated into several peaks due to different molecular weights. The resulting electrophoregram is quite different from that of either CIEF or CGE; therefore, more information about the studied Hb sample can be obtained.
Yang
Chun
C
National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011, China.
Liu
Hechun
H
Yang
Qing
Q
Zhang
Lingyi
L
Zhang
Weibing
W
Zhang
Yukui
Y
D000818:Animals; D002417:Cattle; D003956:Dialysis; D019075:Electrophoresis, Capillary / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D014644:Genetic Variation; D006454:Hemoglobins / Q000235:genetics / Q000302:isolation & purification; D007525:Isoelectric Focusing / Q000379:methods*; D009862:Online Systems; D011506:Proteins / Q000302:isolation & purification; D011817:Rabbits
D006454:Hemoglobins; D011506:Proteins