Effective separation of peptides using highly dense thermo-responsive polymer brush-grafted porous polystyrene beads

Aya Mizutani, Kenichi Nagase, Akihiko Kikuchi, Hideko Kanazawa, Yoshikatsu Akiyama, Jun Kobayashi, Masahiko Annaka, Teruo Okano

Research output: Contribution to journalArticlepeer-review

36 Citations (Scopus)


For the development of well-defined highly dense thermo-responsive polymer grafted surface as an improved stationary phase for thermo-responsive chromatography, poly(N-isopropylacrylamide) (PIPAAm) brush-grafted porous polystyrene beads were prepared by surface-initiated atom transfer radical polymerization (ATRP). The PIPAAm grafted region of polystyrene beads was adjusted by the addition of isooctane as a poor solvent for polystyrene upon the reaction of ATRP initiator immobilization. Using a thermo-responsive HPLC column containing the prepared beads with PIPAAm brush grafted on the inside pores nearby the outer surfaces, angiotensin subtypes were effectively separated with aqueous mobile phase, because the densely grafted PIPAAm on nearby the outer surface effectively interacted with the peptides hydrophobically. Retention of basic peptide was achieved by the beads with basic mobile phase. These results indicated that the prepared beads with grafted PIPAAm nearby the outer surface became an effective chromatographic stationary phase for retaining basic peptides using wide pH range of mobile phase.

Original languageEnglish
Pages (from-to)2191-2198
Number of pages8
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Issue number24
Publication statusPublished - 2010 Aug
Externally publishedYes


  • Atom transfer radical polymerization
  • Peptide
  • Poly(N-isopropylacrylamide)
  • Poly(styrene-co-divinylbenzene)
  • Polymer brush
  • Thermo-responsive chromatography
  • Thermo-responsive polymer

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology


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