Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

J Pharm Biomed Anal. 2005 Jul 1;38(3):408-13. doi: 10.1016/j.jpba.2005.01.019. Epub 2005 Feb 17.

Abstract

A method based on capillary zone electrophoresis (CZE) was used to study the interaction between low molecular weight heparin (LMWH) and interleukin 2 (IL-2). The results showed that the increase of the concentration of LMWH led to the decrease of the peak height and the increase of the peak width of IL-2, but the peak areas were kept constant. The binding constant of IL-2 with LMWH was calculated as 1.2 x 10(6)M(-1) by Scatchard analysis, which is in good agreement with the results found in the references using enzyme-linked immunosorbent assay (ELISA). The results demonstrated that the interaction between IL-2 and LMWH is of fast on-and-off kinetic binding reaction. CZE might be used to study not only slow on-and-off rates interactions, but also fast on-and-off rates ones. The binding constant can be calculated easily, and the method can be applied to study a wide range of heparin-protein interactions.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzoic Acid / chemistry
  • Binding, Competitive
  • Electrophoresis, Capillary / methods*
  • Epidermal Growth Factor / chemistry
  • Heparin, Low-Molecular-Weight / analysis*
  • Heparin, Low-Molecular-Weight / chemistry
  • Humans
  • Interleukin-2 / analysis*
  • Interleukin-2 / chemistry
  • Interleukin-2 / genetics
  • Recombinant Proteins / analysis
  • Recombinant Proteins / chemistry
  • Technology, Pharmaceutical / methods

Substances

  • Heparin, Low-Molecular-Weight
  • Interleukin-2
  • Recombinant Proteins
  • Epidermal Growth Factor
  • Benzoic Acid