The analysis of heparin-protein interactions using evanescent wave biosensor with regioselectively desulfated heparins as the ligands.
Evanescent wave biosensor has been recently employed as a powerful tool for analyses of macromolecular interactions. In the present study, evanescent wave biosensor analysis was developed to analyze the heparin-protein interaction using as ligands a series of heparin derivatives regioselectively desulfated by chemical methods, particularly to evaluate the effect of each sulfate group of heparin. The method for immobilizing heparin on the cuvette of the evanescent wave biosensor equipment was optimized to obtain the high response required for accurate measurement. The best result was achieved when the amino group introduced at the reducing end of heparin was coupled with carboxymethyl dextran on the surface of the cuvette using glycolchitosan as a multivalent linker. The established system appeared to describe well the interactions of heparin with such proteins as acidic and basic fibroblast growth factors and tissue factor pathway inhibitor.
Kamei
K
K
Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan. kame@ipc.kit.ac.jp
Wu
X
X
Xu
X
X
Minami
K
K
Huy
N T
NT
Takano
R
R
Kato
H
H
Hara
S
S
D015374:Biosensing Techniques; D002686:Chitin / Q000031:analogs & derivatives; D048271:Chitosan; D003911:Dextrans; D016220:Fibroblast Growth Factor 1 / Q000737:chemistry; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry; D006493:Heparin / Q000737:chemistry*; D007700:Kinetics; D008024:Ligands; D008074:Lipoproteins / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*
D003911:Dextrans; D008024:Ligands; D008074:Lipoproteins; D011506:Proteins; C051928:lipoprotein-associated coagulation inhibitor; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D002686:Chitin; D006493:Heparin; D048271:Chitosan; C014392:carboxymethyl dextran