Capillary zone electrophoresis for the study of the binding of antithrombin to low-affinity heparin.
When low-affinity interactions between glycosaminoglycans and precious proteins are studied, it is imperative to design an experimental set-up that consumes as little material as possible. To evaluate the applicability of the CZE technique to this problem, we explored the interaction between antithrombin and low-affinity heparin. In a series of CZE experiments we demonstrated that the mobility of antithrombin increases gradually as increased concentrations of low-affinity heparin were added to the electrolyte. The results were, as expected, consistent with the general algorithm for monovalent binding. The binding constant was estimated at 20+/-6 microM in excellent agreement with the value reported in the literature.
Gunnarsson
K
K
Department of Medical Chemistry, Uppsala University, Biomedical Centre, Sweden. Kerstin.K.Gunnarsson@eu.biacore.com
Valtcheva
L
L
Hjertén
S
S
D000818:Animals; D000991:Antithrombins / Q000378:metabolism*; D001665:Binding Sites; D019075:Electrophoresis, Capillary / Q000379:methods*; D006493:Heparin / Q000378:metabolism*; D013552:Swine
D000991:Antithrombins; D006493:Heparin