Temperature-responsive chromatography for the separation of biomolecules

Hideko Kanazawa, Teruo Okano

Research output: Contribution to journalReview articlepeer-review

85 Citations (Scopus)


Temperature-responsive chromatography for the separation of biomolecules utilizing poly (N-isopropylacrylamide) (PNIPAAm) and its copolymer-modified stationary phase is performed with an aqueous mobile phase without using organic solvent. The surface properties and function of the stationary phase are controlled by external temperature changes without changing the mobile-phase composition. This analytical system is based on nonspecific adsorption by the reversible transition of a hydrophilic-hydrophobic PNIPAAm-grafted surface. The driving force for retention is hydrophobic interaction between the solute molecules and the hydrophobized polymer chains on the stationary phase surface. The separation of the biomolecules, such as nucleotides and proteins was achieved by a dual temperature- and pH-responsive chromatography system. The electrostatic and hydrophobic interactions could be modulated simultaneously with the temperature in an aqueous mobile phase, thus the separation system would have potential applications in the separation of biomolecules. Additionally, chromatographic matrices prepared by a surface-initiated atom transfer radical polymerization (ATRP) exhibit a strong interaction with analytes, because the polymerization procedure forms a densely packed polymer, called a polymer brush, on the surfaces. The copolymer brush grafted surfaces prepared by ATRP was an effective tool for separating basic biomolecules by modulating the electrostatic and hydrophobic interactions. Applications of thermally responsive columns for the separations of biomolecules are reviewed here.

Original languageEnglish
Pages (from-to)8738-8747
Number of pages10
JournalJournal of Chromatography A
Issue number49
Publication statusPublished - 2011 Dec 9


  • Oligonucleotides
  • PH- and temperature-responsive polymer
  • Peptide
  • Poly(N-isopropylacrylamide)
  • Protein
  • Temperature-responsive chromatography

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry


Dive into the research topics of 'Temperature-responsive chromatography for the separation of biomolecules'. Together they form a unique fingerprint.

Cite this