Determination of the binding parameters for antithrombin-heparin fragment systems by affinity and frontal analysis continuous capillary electrophoresis.
The suitability of affinity capillary electrophoresis (ACE) and frontal analysis continuous capillary electrophoresis (FACCE) for binding constant determination was investigated for complexes between heparin fragments and antithrombin III, one of the main target proteins in the coagulation cascade. In a 100 mM ionic strength phosphate buffer (pH 7.4), ACE was suitable to determine weak to medium interactions developed by short oligomeric heparin fragments, but it failed for decasaccharide, which presents a more complex irreversible interaction. However FACCE allowed evaluating the binding constant for these longer oligomeric fragments. Both experimental approaches were complementary for a wide variety of heparinic fragments.
Le Saux
T
T
Laboratoire d'Electrochimie et Chimie Analytique, UMR CNRS 7575, Ecole Nationale Supérieure de Chimie de Paris, Université ParisVI, 11, rue Pierre et Marie Curie, 75231 Paris Cedex 05, France.
Varenne
A
A
Perreau
F
F
Siret
L
L
Duteil
S
S
Duhau
L
L
Gareil
P
P
D000991:Antithrombins / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006493:Heparin / Q000737:chemistry*; D008962:Models, Theoretical; D011485:Protein Binding
D000991:Antithrombins; D006493:Heparin