Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2.
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.
Liang
Aiye
A
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China.
He
Xinya
X
Du
Yuguang
Y
Wang
Keyi
K
Fung
Yingsing
Y
Lin
Bingcheng
B
D019817:Benzoic Acid / Q000737:chemistry; D001667:Binding, Competitive; D019075:Electrophoresis, Capillary / Q000379:methods*; D004815:Epidermal Growth Factor / Q000737:chemistry; D006495:Heparin, Low-Molecular-Weight / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D007376:Interleukin-2 / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry; D013678:Technology, Pharmaceutical / Q000379:methods
D006495:Heparin, Low-Molecular-Weight; D007376:Interleukin-2; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D019817:Benzoic Acid