Fast preparation of monolithic immobilized pH gradient column by photopolymerization and photografting techniques for isoelectric focusing separation of proteins

Authors

  • Yu Liang,

    1. Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
    2. Graduate School of the Chinese Academy of Sciences, Beijing, China
    Search for more papers by this author
  • Guijie Zhu,

    1. Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
    2. Graduate School of the Chinese Academy of Sciences, Beijing, China
    Search for more papers by this author
  • Tingting Wang,

    1. Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
    2. Graduate School of the Chinese Academy of Sciences, Beijing, China
    Search for more papers by this author
  • Xiaodan Zhang,

    1. Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
    Search for more papers by this author
  • Zhen Liang,

    1. Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
    Search for more papers by this author
  • Lihua Zhang,

    1. Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
    Search for more papers by this author
  • Yukui Zhang

    Corresponding author
    1. Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
    • Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China Fax: +86-411-84379560
    Search for more papers by this author

Abstract

A new method was developed to prepare monolithic immobilized pH gradient (M-IPG) columns in UV-transparent fused-silica capillaries by the 5-min photopolymerization of acrylamide and N,N′-methylenebisacrylamide, followed by the 20-min photografting of the focused ampholine-derived glycidylmethacrylate monomer on the monolithic matrix, by which the preparation time was reduced, and the stability of the formed pH gradient was improved, compared with our previous methods. Using the prepared M-IPG column, the baseline separation of proteins was achieved according to their pIs. Without carrier ampholytes added in the running buffer, the separated components could be detected with high sensitivity by UV at low wavelength.

Ancillary