On-column capture of a specific protein in capillary electrophoresis using magnetic beads.
A method for capturing specific molecules separated by CE has been explored. To demonstrate on-column capture of migrating analyte molecules, two detection windows were fabricated on a capillary. Magnetic beads containing immobilized molecules that react with the specific molecules under study were placed between the detection windows in the capillary using magnets. Molecules in a sample solution injected into the capillary were separated and detected at the first detection window. After passing through the first detection window, the separated molecules encountered the magnetic beads, where the specific analyte was captured. As a result, the peak area for those analyte molecules decreased or disappeared completely at the second detection window. Rabbit IgG and carbonic anhydrase were employed to demonstrate on-column capture of a specific molecule. For rabbit IgG, magnetic beads containing the immobilized antibody (anti-rabbit IgG) were used. Rabbit IgG molecules were captured on the magnetic beads during CE migration. Furthermore, the capture of carbonic anhydrase was demonstrated by the reaction between magnetic beads (containing immobilized anti-rabbit IgG) and anti-carbonic anhydrase (rabbit IgG), before the beads were packed in the capillary. After packing the magnetic beads in the capillary, a mixture of two proteins was injected into the capillary. Two proteins were detected at the first detection window, while the peak corresponding to carbonic anhydrase disappeared at the second detection window. The results show that using an appropriate antibody, the present technique would be applicable to any proteins.
Kaneta
Takashi
T
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Motooka, Fukuoka, Japan.
Inoue
Junji
J
Koizumi
Mototsugu
M
Imasaka
Totaro
T
D000818:Animals; D002256:Carbonic Anhydrases / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D052203:Ferrosoferric Oxide / Q000737:chemistry*; D007074:Immunoglobulin G / Q000302:isolation & purification; D008863:Microspheres; D011506:Proteins / Q000302:isolation & purification*; D011817:Rabbits
D007074:Immunoglobulin G; D011506:Proteins; D002256:Carbonic Anhydrases; D052203:Ferrosoferric Oxide