On-column capture of a specific protein in capillary electrophoresis using magnetic beads

Takashi Kaneta, Junji Inoue, Mototsugu Koizumi, Totaro Imasaka

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)3218-3223
Number of pages6
JournalElectrophoresis
Volume27
Issue number16
DOIs
Publication statusPublished - Aug 2006
Externally publishedYes

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Keywords

  • CE
  • Immunoaffinity
  • Magnetic beads
  • Protein

ASJC Scopus subject areas

  • Clinical Biochemistry

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