Interactions of antithrombin and proteins in the plasma contact activation system with immobilized functional heparin.
The interactions of antithrombin (AT) and the contact phase clotting factors with two commercially available heparinized surfaces are reported. The Carmeda (CBAS) and Corline surfaces along with controls (a sulfonated polyethylene surface and a CBAS analog in which the heparin used was devoid of specific AT-binding sequences) were exposed to human plasma. Adsorbed proteins were eluted and examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The CBAS and Corline surfaces adsorbed large amounts of AT, whereas adsorption on the controls was negligible. Immunoblots for the four contact phase clotting factors indicated less contact activation on the CBAS and Corline surfaces than on the controls. Determination of adsorbed functional AT using a FXa inhibition assay showed that the CBAS surface adsorbed about 4 times as much AT as the Corline surface. Adsorption of AT to the control surfaces was minimal. Assays for adsorbed FXII and FXIIa based on kallikrein generation showed that all four surfaces adsorbed similar amounts of FXII. However, on the controls, most of the FXII was in activated form, whereas on the CBAS and Corline surfaces very little activation occurred.
Cornelius
Rena M
RM
Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada L8S 4L7.
Sanchez
Javier
J
Olsson
Per
P
Brash
John L
JL
D000991:Antithrombins / Q000378:metabolism*; D001672:Biocompatible Materials / Q000378:metabolism*; D001779:Blood Coagulation Factors / Q000378:metabolism*; D005172:Factor XI / Q000378:metabolism; D005174:Factor XII / Q000378:metabolism; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D007704:Kininogens / Q000378:metabolism; D010949:Plasma / Q000378:metabolism; D011288:Prekallikrein / Q000378:metabolism
D000991:Antithrombins; D001672:Biocompatible Materials; D001779:Blood Coagulation Factors; D007704:Kininogens; D005174:Factor XII; D006493:Heparin; D005172:Factor XI; D011288:Prekallikrein