Aqueous chromatographic system for separation of biomolecules using thermoresponsive polymer modified stationary phase

Hideko Kanazawa, Mayumi Nishikawa, Aya Mizutani, Chikako Sakamoto, Yuko Morita-Murase, Yoshiko Itoh, Akihiko Kikuchi, Teruo Okano

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

We have investigated a new method for HPLC using packing materials modified with a functional polymer, such as thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm). PNIPAAm-modified silica exhibits temperature-controlled hydrophilic-hydrophobic surface property changes in aqueous systems. Temperature-responsive chromatography is performed with an aqueous mobile phase without using an organic solvent. We designed ternary copolymers of NIPAAm introduced 2-(dimethyl-amino) ethyl methacrylate (DMAEMA) as a cationic monomer and butyl methacrylate (BMA) as a hydrophobic monomer. A cationic thermoresponsive hydrogel grafted surface would produce an alterable stationary phase with both thermally regulated hydrophobicity and charge density for separation of bioactive compounds. In this study, we achieved successful separation of lysozyme without the loss of bioactivity by temperature-responsive chromatography. 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.

Original languageEnglish
Pages (from-to)157-161
Number of pages5
JournalJournal of Chromatography A
Volume1191
Issue number1-2
DOIs
Publication statusPublished - 2008 May 16

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Biomolecules
Polymers
Temperature
Chromatography
Hydrophobic and Hydrophilic Interactions
Functional polymers
Methacrylates
Surface Properties
Hydrogel
Hydrophobicity
Muramidase
Bioactivity
Charge density
Static Electricity
Silicon Dioxide
Organic solvents
Surface properties
Electrostatics
Copolymers
Monomers

Keywords

  • Human serum albumin
  • Lysozyme
  • Peptide
  • Poly(N-isopropylacrylamide) (PNIPAAm)
  • Protein
  • Temperature-responsive chromatography

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Aqueous chromatographic system for separation of biomolecules using thermoresponsive polymer modified stationary phase. / Kanazawa, Hideko; Nishikawa, Mayumi; Mizutani, Aya; Sakamoto, Chikako; Morita-Murase, Yuko; Itoh, Yoshiko; Kikuchi, Akihiko; Okano, Teruo.

In: Journal of Chromatography A, Vol. 1191, No. 1-2, 16.05.2008, p. 157-161.

Research output: Contribution to journalArticle

Kanazawa, Hideko ; Nishikawa, Mayumi ; Mizutani, Aya ; Sakamoto, Chikako ; Morita-Murase, Yuko ; Itoh, Yoshiko ; Kikuchi, Akihiko ; Okano, Teruo. / Aqueous chromatographic system for separation of biomolecules using thermoresponsive polymer modified stationary phase. In: Journal of Chromatography A. 2008 ; Vol. 1191, No. 1-2. pp. 157-161.
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