Analysis of peptides and proteins by temperature-responsive chromatographic system using N-isopropylacrylamide polymer-modified columns

Hideko Kanazawa, Kazuo Yamamoto, Yuki Kashiwase, Yoshikazu Matsushima, Nobuharu Takai, Akihiko Kikuchi, Yasuhisa Sakurai, Teruo Okano

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

A new method of HPLC using packing materials modified with a temperature responsive polymer, poly(N-isopropylacrylamide) (PIPAAm), was developed. Homogeneous PIPAAm polymer and its copolymer with butyl methacrylate (BMA) were synthesized and grafted to aminopropyl silica by activated ester-amine coupling and they were used as packing materials. The surface properties and functions of the stationary phases are controlled by external temperature. Isocratic elution by aqueous mobile phase alone is the basis for separation of peptides and protein. The separation of the mixture of three peptides, insulin chain A and B and β-endorphin fragment 1-27 was achieved by changing the column temperature with 0.9% NaCl aqueous solution as the sole eluent. Retention of peptides and proteins was controlled both by column temperature and by NaCl concentration in the aqueous mobile phases in this chromatographic system.

Original languageEnglish
Pages (from-to)1545-1550
Number of pages6
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume15
Issue number9-10
DOIs
Publication statusPublished - 1997 Jun

Fingerprint

Polymers
Peptides
Temperature
Proteins
Insulin
Endorphins
Surface Properties
Silicon Dioxide
Amines
Surface properties
Esters
Copolymers
High Pressure Liquid Chromatography
N-isopropylacrylamide
poly-N-isopropylacrylamide

Keywords

  • Peptide
  • Poly(N-isopropylacrylamide)
  • Protein
  • Temperature-responsive chromatography

ASJC Scopus subject areas

  • Analytical Chemistry
  • Pharmaceutical Science

Cite this

Analysis of peptides and proteins by temperature-responsive chromatographic system using N-isopropylacrylamide polymer-modified columns. / Kanazawa, Hideko; Yamamoto, Kazuo; Kashiwase, Yuki; Matsushima, Yoshikazu; Takai, Nobuharu; Kikuchi, Akihiko; Sakurai, Yasuhisa; Okano, Teruo.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 15, No. 9-10, 06.1997, p. 1545-1550.

Research output: Contribution to journalArticle

Kanazawa, Hideko ; Yamamoto, Kazuo ; Kashiwase, Yuki ; Matsushima, Yoshikazu ; Takai, Nobuharu ; Kikuchi, Akihiko ; Sakurai, Yasuhisa ; Okano, Teruo. / Analysis of peptides and proteins by temperature-responsive chromatographic system using N-isopropylacrylamide polymer-modified columns. In: Journal of Pharmaceutical and Biomedical Analysis. 1997 ; Vol. 15, No. 9-10. pp. 1545-1550.
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