RSS-Feed abonnieren
DOI: 10.1055/s-0038-1647037
A Simple Method to Measure Dermatan Sulfate at Sub-Microgram Concentrations in Plasma
Publikationsverlauf
Received 30. März 1988
Accepted after revision 07. Juni 1988
Publikationsdatum:
28. Juni 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/198802/lookinside/thumbnails/10-1055-s-0038-1647037-1.jpg)
Summary
A simple method for biological assay of dermatan sulfate (DS) in plasma is described. DS accelerates thrombin inhibition by heparin cofactor II (HC II). The principle of the assay is to measure the residual amidolytic thrombin activity after a short period of incubation with HC II in defibrinated plasma at low ionic strength. For this method we take advantage of two observations. Firstly, at fixed concentrations of DS and of HC II, the rate of thrombin inhibition increases when the ionic strength of the medium decreases. Secondly, defibrination by bentonite absorption also removes antithrombin III, HC II and for a large part alpha-2 macroglobulin from the plasma, so that no other thrombin inhibitor competes with HC II added as a reagent in a second step.
In the conditions described, there is a linear relationship between DS concentrations in plasma from 0 to 2 μg/ml and the log of residual thrombin activity. The limit of sensitivity is 0.1 μg/ ml. The assay displays an acceptable reproducibility in intraassay, inter-assay and inter-individual experiments. It can be used to measure DS in human, rabbit and rat plasmas. The assay is also sensitive to other HC II activators such as heparin and pentosan polysulfate.
DS is effective in experimental thrombosis without any detectable anticoagulant effect ex vivo. Pharmacological concentrations of DS in plasma fall into the range of sensitivity of this assay, which would be helpful in experimental or clinical studies of DS and related glycosaminoglycans.
-
References
- 1 Lindahl U, Hook M. Glycosaminoglycans and their binding to biological macromolecules.. Annu Rev Biochem 1978; 47: 385-417
- 2 Tollefsen DM, Petska CA, Monafo WJ. Activation of heparin cofactor II by dermatan sulfate.. J Biol Chem 1983; 258: 6713-6716
- 3 Teien AN, Abildgaard U, Hook M. The anticoagulant effect of heparan sulfate and dermatan sulfate.. Thromb Res 1976; 8: 859-867
- 4 Sié P, Ofosu F, Fernandez F, Buchanan MR, Petitou M, Boneu B. Respective role of antithrombin III and heparin cofactor II in the in vitro anticoagulant effect of heparin and of various sulphated polysaccharides.. Br J Haematol 1986; 64: 707-714
- 5 Buchanan MR, Boneu B, Ofosu F, Hirsh J. The relative importance of thrombin inhibition and factor Xa inhibition to the antithrombotic effects of heparin.. Blood 1985; 65: 198-201
- 6 Fernandez F, Van RynJ, Ofosu F, Hirsh J, Buchanan MR. The haemorragic and antithrombotic effects of dermatan sulphate.. Br J Haematol 1986; 64: 309-317
- 7 Dol F, Sie P, Dupouy D, Boneu B. Effect of pentosan polysulphate administration in man on the formation of covalent complexes between heparin cofactor II and thrombin generated in plasma by contact activation.. Thromb Haemostas 1986; 56: 295-298
- 8 Sie P, Lansen J, Lacheretz F, Verschuere B, Boneu B. Comparative turn-over of heparin cofactor II and antithrombin III in baboons.. Thromb Haemostas 1986; 56: 302-307
- 9 Sie P, Dupouy D, Pichon J, Boneu B. Constitutional heparin cofactor II deficiency associated with recurrent thrombosis.. Lancet 1985; II: 414-416
- 10 Tollefsen DM, Majerus DW, Blank MK. Heparin cofactor II. Purification and properties of a heparin-dependent inhibitor of thrombin in human plasma.. J Biol Chem 1982; 257: 2162-2169
- 11 Dawes J, Pepper DS. A sensitive competitive binding assay for exogenous and endogenous heparin.. Thromb Res 1982; 27: 387-396
- 12 Wunderwald P, Schrenk WJ, Port H. Antithrombin BM from human plasma: an antithrombin binding moderately to heparin.. Thromb Res 1982; 25: 177-191
- 13 Scully MF, Ellis V, Kakkar VV. Comparison of the molecular mass dependency of heparin stimulation of heparin cofactor II-thrombin interaction to antithrombin Ill-thrombin interaction.. Thromb Res 1987; 46: 491-502
- 14 Tollefsen DM, Petska CA. Modulation of heparin cofactor II activity by histidine rich glycoprotein and platelet factor 4.. J Clin Invest 1985; 75: 496-501
- 15 Preissner KT, Sie P. S protein-vitronectin neutralizes the anticoagulant activity of glycosaminoglycans in the inhibition of thrombin by heparin cofactor II.. Thromb Haemostas 1987; 58: 227 (Abstr)
- 16 Hamazaki H. Ca2+-mediated association of human serum amyloid P component with heparan sulfate and dermatan sulfate.. J Biol Chem 1987; 262: 1456-1460
- 17 Handin RI, Cohen HJ. Purification and binding properties of human platelet factor four.. J Biol Chem 1976; 251: 4273-4282
- 18 Griffith MJ, Marbet GA. Dermatan sulfate and heparin can be fractionated by affinity for heparin cofactor II.. Biochem Biophys Res Commun 1983; 112: 663-670