RSS-Feed abonnieren
DOI: 10.1055/s-0038-1657891
Quantitative Assessment of Soluble Fibrin in Plasma by Affinity Chromatography – A Comparative Study with desAA-Fibrin, desAABB-Fibrin and Fibrinogen
Publikationsverlauf
Received 27. März 1985
Accepted 20. Juni 1985
Publikationsdatum:
18. Juli 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/198502/lookinside/thumbnails/10-1055-s-0038-1657891-1.jpg)
Summary
A quantitative determination of soluble fibrin in plasma was carried out by affinity chromatography. For this purpose, desAA-fibrin and fibrinogen immobilized on Sepharose 4B were used at the stationary side whereas batroxobin-induced 125I-desAA-fibrin or thrombin-induced 125I-desAABB-fibrin mixed with plasma containing 131I-fibrinogen represented the fluid phase. The binding characteristics of these mixtures to the immobilized proteins were compared at 20° C and 37° C. Complete binding of both types of fibrin to the immobilized desAA-fibrin was always seen at 20° C as well as at 37° C. However, binding of soluble fibrin was accompanied by substantial binding of fibrinogen that was more pronounced at 20° C. Striking differences depending on the temperature at which the affinity chromatography was carried out, were documented for the fibrinogen-fibrin interaction. At 20° C more than 90% of the applied desAA-fibrin was bound to the immobilized fibrinogen whereas at 37° C only a mean of 17% were retained at the fibrinogen-Sepharose column. An opposite finding with regard to the tested temperature was made with the desAABB-fibrin. Nearly complete binding to insolubilized fibrinogen was found at 37° C (95%) but only 58% of the desAABB-fibrin were bound at 20° C. The binding patterns did not change when the experiments were performed in the presence of calcium ions. The opposite behaviour of the two types of soluble fibrin to immobilized fibrinogen at the different temperatures, together with the substantial binding of fibrinogen in the presence of soluble fibrin to insolubilized fibrin in every setting tested, devaluates affinity chromatography as a tool in the quantitative assessment of soluble fibrin in patients’ plasma.
-
References
- 1 Blombäck B, Hessel B, Hogg D, Therkildsen L. A two-step fibrinogen-fibrin transition in blood coagulation. Nature 1978; 275: 501-505
- 2 Apitz K. Über Profibrin. I. Die Entstehung und Bedeutung des Profibrins im Gerinnungsverlauf. Zschr Ges Exp Med 1937; 101: 552-584
- 3 Thomas L, Smith RT, von Korff RT. Cold-precipitation by heparin of a protein in rabbit and human plasma. Proc Soc Exp Biol Med 1954; 86: 813-818
- 4 Shainoff JR, Page IH. Significance of cryoprofibrin in fibrinogen-fibrin conversion. J Exp Med 1962; 116: 687-707
- 5 Heene DL, Matthias FR. Adsorption of fibrinogen derivates on insolubilized fibrinogen and fibrinmonomer. Thromb Res 1973; 02: 137-154
- 6 Yudelman I, Spanondis K, Nossel HL. 125I-fibrinogen and 125I-fibrin in solution. Thromb Res 1974; 05: 495-509
- 7 Matthias FR, Reinicke R, Heene DL. Affinity chromatography and quantification of soluble fibrin from plasma. Thromb Res 1977; 10: 365-384
- 8 Semeraro N, Collen D. Affinity chromatography of soluble fibrin in human plasma on insolubilized fibrinogen and fragment D. Thromb Res 1978; 12: 1059-1070
- 9 Kierulf P, Brosstad F, Godal HC, Lund PK, Anderson AB, Naeverlid I. Poor discriminating power of fibrinogen-Sepharose towards plasma fibrinogen, fibrin des-AA and fibrin des-AABB monomers, as studied by labelled proteins, laser nephelometric FR-antigen and RIA-FPA quantification. Thromb Res 1979; 16: 01-10
- 10 Sasaki T, Page IH, Shainoff JR. Stable complex of fibrinogen and fibrin. Science 1966; 152: 1069-1071
- 11 Smith GF. Fibrinogen-fibrin conversion. The mechanism of fibrin-polymer formation in solution. Biochem J 1980; 185: 01-11
- 12 Fletcher AP, Alkjaersig N, O’Brain J, Tulevski VG. Blood hypercoagulability and thrombosis. Trans Assoc Am Physicians 1970; 83: 159-166
- 13 von Hugo R, Hafter R, Stemberger A, Graeff H. Soluble fibrin monomer complexes demonstrated by agarose gel filtration and by adsorption on insolubilized fibrinogen. Thrombos Diathes Haemorrh 1975; 34: 216-222
- 14 Donati MB, Verhaeghe R, Culasso DE, Vermylen J. Molecular size distribution of fibrinogen derivates formed in vitro and in vivo: a chromatography study. Thromb Res 1976; 36: 14-26
- 15 Brosstad F. Different chromatographic behaviour of soluble, non-FSF-stabilized des-AA fibrin/fibrinogen and des-AABB fibrin/fibrinogen complexes. Thromb Res 1979; 15: 563-568
- 16 Müller-Berghaus G, Mahn I, Krell W. Formation and dissociation of soluble fibrin complexes in plasma at 20°C and at 37°C. Thromb Res 1979; 14: 561-572
- 17 Kröhnke I, Mahn I, Krell W, Müller-Berghaus G. Studies on circulating soluble fibrin: Separation of 125I-des-AB fibrin and 131I-fibrinogen by gel filtration. Br J Haematol 1980; 45: 131-141
- 18 Vermylen J, Donati MB, Verstraete M. The identification of fibrinogen derivates in plasma and serum by agarose gel filtration. Scand J Haematol 1971; 13: 219-223
- 19 Graeff H, von Hugo R. Identification of fibrinogen derivates in plasma samples. Thrombos Diathes Haemorrh 1972; 27: 610-618
- 20 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Ark Kemi 1956; 10: 415-443
- 21 Mahn I, Müller-Berghaus G. Studies of catabolism of 125I-labelled fibrinogen in normal rabbits and in rabbits with indwelling intravenous catheters: Methodologic aspects. Haemostasis 1975; 04: 40-50
- 22 Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl-sulfate-polyacrylamid gel electrophoresis. J Biol Chem 1969; 24: 4406-4412
- 23 McFarlane AS. Efficient trace-labelling of proteins with iodine. Nature 1958; 182: 53
- 24 Kehl M, Lottspeich F, Henschen A. Analysis of human fibrinopep-tides by high performance-liquid chromatography. Hoppe-Seyler’s Z Physiol Chem 1981; 362: 1661-1664
- 25 Godal HC, Abildgaard U. Gelation of soluble fibrin in plasma by ethanol. Scand J Haemat 1966; 03: 342-350
- 26 Seaman A. Recognition of intravascular clotting. The plasma protamine paracoagulation test. Arch Int Med 1970; 125: 1016-1021
- 27 Tomikawa M, Iwamoto M, Olsson P, Söderman S, Blombäck B. On the platelet-fibrinogen interaction. Thromb Res 1980; 19: 869-876
- 28 Scheefers-Borchel U, Müller-Berghaus G, Fuhge P, Heimburger N. Discrimination between fibrin and fibrinogen by monoclonal antibodies to a synthetic hexapeptide representing the amino-terminus of the alpha-chain of fibrin. Circulation 1984; 70: 11/93 (Abstr.)