Estimation of surface-bound heparin activity: a comparison of methods.
We compared two assays for estimating the amount of active heparin bound to a catheter surface: 1) a kinetic assay based on the inactivation of thrombin by antithrombin III, and 2) thrombin uptake. Both assays were used to estimate the amount of heparin activity on a series of catheters coated with no heparin, covalently bound heparin, and ionically bound heparin. The kinetic assay produced estimates of surface-bound heparin activity and showed that some binding methods resulted in destruction of most of the heparin's biologic activity. In contrast, the thrombin uptake assay did not correlate with the amount of heparin activity on the catheter surface. Substantial thrombin uptake was found on surfaces coated with no heparin or inactive heparin, while low thrombin uptake was found on surfaces with high levels of heparin activity in the kinetic assay. We conclude that: 1) a kinetic assay based on the heparin accelerated inactivation of thrombin by antithrombin III can be used to estimate the amount of active heparin bound to a catheter surface, and 2) thrombin uptake studies do not correlate with heparin activity and do not predict which heparin binding method will result in the highest concentration of active heparin on the catheter surface.
Chandler
W L
WL
Department of Laboratory Medicine, University of Washington, Seattle 98195.
Solomon
D D
DD
Hu
C B
CB
Schmer
G
G
D000327:Adsorption; D002404:Catheterization; D006493:Heparin / Q000032:analysis / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D013053:Spectrophotometry / Q000379:methods; D013917:Thrombin / Q000378:metabolism
D006493:Heparin; D013917:Thrombin