Retention and
Thermodynamic Properties of Three Insulin Variants on
the Reversed-phase Liquid Chromatography
GUO Min-Liang1*, Milton T W HEARN2,
Reinhard I BOYSEN2
(1College of Bioscience and Biotechnology, Yangzhou University,
Jiangsu 225009, China; 2Centre for Bioprocess
Technology, Department of Biochemistry and Molecular Biology, Monash University,
Clayton, Victoria 3168, Australia )
Abstract The retention
and thermodynamic behaviour of three insulin variants, bovine, human and
porcine insulins, in reversed-phase high-performance liquid chromatography were
studied over a range of temperatures between 10 and 65 ℃ and a range of
methanol concentrations between 53 and 59%(v/v). The results demonstrated
that the slight difference of three insulins in the amino acid sequence could
be resolved significantly in C8-hydrophobic ligand. Values for the
relative changes in enthalpy (ΔHo/R) and entropy (ΔSo*) associated
with the interaction process were also determined. These values of
thermodynamic parameters would provide further insight into the factors
involved in the stabilization of protein conformation and the mechanism of the
interaction of peptides with hydrophobic surfaces. The experimental results
also demonstrated that the determination of thermodynamic parameters of
interactions between peptides and hydrophobic surfaces would provide an
alternative approach for the investigation of mechanisms of protein folding and
of interaction between proteins.
Key words reversed-phase
liquid chromatography； peptide and
protein； retention behaviour； thermodynamic parameter
Received:
November 8, 1999 Accepted: December 30, 1999
* Corresponding author: Tel, 86-514-7979056; Fax,
86-514-7979343; e-mail, [email protected]