Soluble Crosslinked Fibrin(ogen) Polymers

Summary

The present study is concerned with the formation of fibrin-fibrinogen associations in the presence of FXIIIa while fibrinolysis was inhibited by aprotinin and EACA. SDS agarose-poly acryl-amide gel electrophoresis on 2.5% gels under non-reducing conditions and ultracentrifugation of the associations on urea-sucrose density gradients showed the formation of soluble crosslinked high molecular weight (HMW) fibrin-fibrinogen polymers with an estimated molecular size up to 10 times that of fibrinogen. After incubation of a mixture of ¹³¹I-fibrinogen (4 mg/ml) and ¹²⁵I-desAA-fibrin (0.2 mg/ml) with FXIIIa (2 U/ml) for 1 h at 37ΰ C, about 5% of the fibrinogen and 80% of the fibrin was incorporated into the generated soluble HMW polymers. The detection of soluble crosslinked fibrin-fibrinogen polymers could be a useful diagnostic criterion for imminent DIC.

Key words

Soluble fibrin - Fibrinogen - FXIIIa - Prevention of clot formation

• References

• 1 Krell W, Mahn I, Heimburger N, Müller-Berghaus G. Fibrin-fibrinogen interactions during gel filtration in a medium containing fibrinogen, albumin or cold-insoluble globulin. Thromb Res 1981; 23: 41-50
• 2 Yudelman I, Spanondis K, Nossel HL. Comparative behaviour of ¹²⁵I-fibrinogen and ¹²⁵I-fibrin in solution. Thromb Res 1974; 5: 495-509
• 3 Selmayr E, Mahn I, Müller-Berghaus G. Crosslinking of soluble fibrin and fibrinogen. Thromb Res 1985; 39: 467-474
• 4 Müller-Berghaus G, Mahn I, Krell W. Formation and dissociation of soluble fibrin complexes in plasma at 20° C and 37° C. Thromb Res 1979; 14: 561-572
• 5 Bale MD, Janmey PA, Ferry JD, Lorand L. Kinetics of formation of fibrin oligomers III. Ligation kinetics concurrent with and subsequent to oligomer assembly. Biopolymer 1984; 23: 127-138
• 6 Preissner KT, Rötker J, Selmayr E, Fasold H, Müller-Berghaus G. Influence of fibrinogen on fibrin polymerization. Ultracentrifugation studies. Biochim Biophys Acta 1985; 829: 358-364
• 7 Sasaki T, Page I, Shainoff JR. Stable complex of fibrinogen and fibrin. Science 1966; 152: 1069-1071
• 8 Kanaide H, Shainoff JR. Crosslinking of fibrinogen and fibrin by fibrin-stabilizing-factor (factor XHIa). J Lab Clin Med 1975; 85: 574-579
• 9 Furlan M, Seelich T, Beck EA. Clottability and crosslinking reactivity of fibrin(ogen) following differential release of fibrinopeptides A and B. Thromb Haemostas 1976; 36: 582-592
• 10 Ly B, Kierulf P, Jacobsen E. Stabilisation of soluble fibrin/fibrinogen complexes by fibrin-stabilizing factor (FSF). Thromb Res 1974; 4: 509-522
• 11 Kierulf P, Brosstad F, Andersen AB. FSF-stabilized soluble fibrinogen/fibrin intermediates in “Ca++-plasma” at 37° C. Scand J Haematol 1983; 30: 97-102
• 12 Sherman LA, Harwig S, Lee S. In vitro formation and in vivo clearance of fibrinogen-fibrin complexes. J Lab Clin Med 1975; 86: 100-111
• 13 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Ark Kemi 1956; 10: 415-443
• 14 Mahn I, Müller-Berghaus G. Studies on catabolism of ¹²⁵-I-labelled fibrinogen in normal rabbits with indwelling intravenous catheter: Methodologic aspects. Haemostasis 1975; 4: 40-50
• 15 Brown ME, Rothstein F. Fibrinogen from human plasma: Preparation by precipitation with heavy-metal coordination complex. Science 1967; 19: 1017-1019
• 16 McFarlane AS. Efficient tracer-labelling of proteins with iodine. Nature 1958; 128: 53
• 17 Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl-sulfate-polyacrylamid gel electrophoresis. J Biol Chem 1969; 24: 4406-4412