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DOI: 10.1160/TH09-03-0168
Detection and neutralisation of heparin by a fluorescent ruthenium compound
Financial support: This work was supported by the European Commission (Marie Curie Intra-European Fellowship for H. Szelke).Publikationsverlauf
Received:
16. März 2009
Accepted after major revision:
16. Juni 2009
Publikationsdatum:
27. November 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/200905/lookinside/thumbnails/10-1160-th09-03-0168-1.jpg)
Summary
Heparin and low-molecular-weight heparin (LMWH) are commonly monitored by determination of activated clotting times or chromogenic assays. Despite their wide use, these assays determine the biological activity and not the concentration of the anticoagulants. They may be inaccurate in some circumstances such as certain disease states. In addition, there is a significant interest in alternative tests for the point-of-care detection of heparin and LMWH. Their binding to small molecules for the detection in biological matrices is poorly explored. We describe here a new optical molecular probe for the detection of LMWH in serum samples. The polycationic ruthenium compound 1 is applicable to the quantification of heparin by monitoring 630 nm fluorescence. In addition, compound 1 is a rare example of a non-polymeric low molecular weight compound which neutralises the anticoagulant activity of heparin and LMWH in plasma samples. Limitation of the method is its low sensitivity currently being improved by structural modification of compound 1.
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References
- 1 Van Kerkhof J C, Bergveld P, Schasfoort R B M. The ISFET based heparin sensor with a monolayer of protamine as affinity ligand. Biosens Bioelectron 1995; 10: 269-282.
- 2 Despotis G J, Gravlee G, Filos K. et al. Anticoagulation monitoring during cardiac surgery: A review of current and emerging techniques. Anesthesiology 1999; 91: 1122-1151.
- 3 Jelinek R, Kolusheva S. Carbohydrate Biosensors. Chem Rev 2004; 104: 5987-6015.
- 4 Wright Z LZhong, Anslyn E V. A functional assay for heparin in serum using a designed synthetic receptor. Angew Chem Int Ed 2005; 44: 5679-682.
- 5 Wang S, Chang Y-T. Discovery of heparin chemosensors through diversity oriented fluorescence library approach. Chem Commun 2008; 1173-1175.
- 6 Metz S, Horrow J C. Pharmacology & Physiology in Anesthetic Practice. ed. Stoelting RK. Lippincott J B. Co; Philadelphia, PA: 1994: 1-15.
- 7 Verrecchio A, Germann M W, Schick B P. et al. Design of peptides with high affinities for heparin and endothelial cell proteoglycans. J Biol Chem 2000; 275: 7701-7707.
- 8 Schick B P, Gradowski J F, San J DAntonio. et al. Novel design of peptides to reverse the anticoagulant activities of heparin and other glycosaminoglycans. Thromb Haemostas 2001; 85: 482-487.
- 9 Fromm J R, Hileman R E, Caldwell E E O. et al. Pattern and spacing of basic amino acids in heparin binding sites. Arch Biochem Biophys 199; 343: 92-100.
- 10 Potempa L A, Gewurz H. Influence of heparin on interactions between C-reactive protein and polycations. Mol Immunol 1983; 20: 501-509.
- 11 Chang L-C, Liang J F, Lee H-F. et al. Low molecular weight protamine (LMWP) as nontoxic heparin/low molecular weight heparin antidote (II): in vitro evaluation of efficacy and toxicity. AAPS pharmSci 2001; 03: E18.
- 12 Choi S, Clements D J, Pophristic V. et al. The design and evaluation of heparin-binding foldamers. Angew Chem Int Ed 2005; 44: 6685-6689.
- 13 Wu H-f, Lundblad R L, Church F C. Neutralization of heparin activity by neutrophil lactoferrin. Blood 1995; 85: 421-428.
- 14 Fabian I, Aronson M. Polycations as possible substitutes for protamine in heparin neutralization. Thromb Res 1980; 17: 239-247.
- 15 van deWesterlo E M A, Smetsers T F C M, Dennissen M A B A. et al. Human single chain antibodies against heparin: selection, characterization, and effect on coagulation. Blood 2002; 99: 2427-2433.
- 16 Croether M A, Warkentin T E. Bleeding risk and the management of bleeding complications in patients undergoing anticoagulant therapy: focus on new anticoagulants. Blood 2008; 111: 4871-4879.
- 17 Mecca T, Consoli G M L, Geraci C. et al. Polycationic calyx[8]arenes able to recognize and neutralize heparin. Org Biomol Chem 2006; 04: 3763-3768.
- 18 Rozenberg G I, Espada J, de Cidre L L. et al. Heparan sulfate, heparin, and heparinase activity detection on polyacrylamide gel electrophoresis using the fluorochrome tris(2,2’-bipyridine)ruthenium(II). Electrophoresis 2001; 22: 3-11.
- 19 Harenberg J, Giese C, Dempfle C E. et al. Monitoring of heparin and low molecular weight heparin with capillary and venous whole blood. Thromb Haemostas 1988; 60: 377-381.
- 20 Creutz C, Sutin N. Electron Transfer Reactions of Excited States: Direct Evidence for Reduction of the Charge Transfer Excited State of Tris(2,2’-bipyridine) ruthenium(II). J Am Chem Soc 1976; 98: 6384-6385.
- 21 Casu B. Structure and active domain of heparin. In: Chemistry and Biology of Heparin and Heparan Sulfate. Oxford: Elsevier; 2005: 1-28.
- 22 Mulloy B, Forster M J, Jones C. et al. NMR and molecular-modelling studies of the solution conformation of heparin. Biochem J 1993; 293: 849-858.
- 23 Conrad H E. Heparin-Binding Proteins. San Diego: Academic Press; 1998