Affinity capillary electrophoresis for the determination of binding affinities for low molecular weight heparins and antithrombin-III

2014 ◽  
Vol 35 (10) ◽  
pp. 1469-1477 ◽  
Author(s):  
Meredith M. Dinges ◽  
Kemal Solakyildirim ◽  
Cynthia K. Larive
Keyword(s):  
Molecular Weight ◽  
Capillary Electrophoresis ◽  
Antithrombin Iii ◽  
Low Molecular Weight ◽  
Binding Affinities ◽  
Low Molecular Weight Heparins ◽  
Affinity Capillary Electrophoresis

The Additive Effect of Low Molecular Weight Heparins on Thrombin Inhibition by Dermatan Sulfate

1993 ◽  
Vol 70 (03) ◽  
pp. 443-447 ◽  
Author(s):  
Benilde Cosmi ◽  
Giancarlo Agnelli ◽  
Edward Young ◽  
Jack Hirsh ◽  
Jeffrey Weitz
Keyword(s):  
Molecular Weight ◽  
Antithrombin Iii ◽  
Dermatan Sulfate ◽  
Anticoagulant Activity ◽  
Additive Effect ◽  
Low Molecular Weight ◽  
Low Molecular Weight Heparins ◽  
Heparin Cofactor Ii ◽  
Thrombin Inhibition ◽  
Sds Page

SummaryThe aim of this study was to investigate the mechanism by which the anticoagulant activity of dermatan sulfate (DS) is increased by low molecular weight heparin (LMWH). In platelet poor plasma, LMWH enhances the effect of DS on thrombin (IIa) inhibition as determined by thrombin clotting times and with a chromogenic substrate assay. Analysis of the results of the chromogenic assays using either the algebraic fractional or the graphic isobole method suggests that LMWH has an additive effect on the anti-IIa activity of DS. This additive effect was lost when the experiments were repeated in plasma immunodepleted of antithrombin III (ATIII), indicating that the anti-IIa activity of LMWH is ATIII-dependent. To further explore the mechanism of the interaction between LMWH and DS, 125I-labeled IIa was added to plasma in the presence or absence of DS and/or LMWH and the formation of IIa-inhibitor complexes was assessed using SDS-PAGE followed by autoradiography. DS addition selectively increases the formation of heparin cofactor II (HCII)-IIa complexes, whereas LMWH enhances ATIII-IIa complex generation. Compared to plasma containing DS alone, the formation of ATIII-IIa complexes also is increased when the combination of DS and LMWH is added. These findings suggest that the additive effect of LMWH on the anti-IIa activity of DS reflects their different modes of IIa inhibition; DS potentiates IIa inhibition by HCII, while LMWH catalyses ATIII-dependent IIa inactivation. The potential clinical significance of these findings requires further investigation.

Determination of Low-Molecular-Weight Heparins and Their Binding to Protamine and a Protamine Analog Using Polyion-Sensitive Membrane Electrodes

1999 ◽  
Vol 266 (1) ◽  
pp. 116-124 ◽  
Author(s):  
Narayanan Ramamurthy ◽  
Narayan Baliga ◽  
Thomas W. Wakefield ◽  
Philip C. Andrews ◽  
Victor C. Yang ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Low Molecular Weight ◽  
Membrane Electrodes ◽  
Low Molecular Weight Heparins ◽  
Sensitive Membrane
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