Determination of Soluble Fibrin Monomer Complexes by Adsorption on Immobilized Fibrinogen

Summary

Determination of fibrin in plasma, also containing fibrinogen, is possible by selective adsorption on affinity columns prepared from highly purified fibrinogen. Desorption is complete using 1.0 M potassium bromide, pH 5.3. In addition, the cold-insoluble globulin is desorbed. Therefore, the eluate is checked on microtiter plates by the staphylococcal clumping test which selectively detects fibrin or fibrinogen. The procedure works well with simple laboratory equipment. The diagnosis of fibrinogen-fibrin complexes is demonstrated in some pathological plasmas.

• References

• 1 AxéN R, Ernback S. 1971; Chemical fixation of enzymes to cyanogen halide activated polysaccharide carriers. European Journal of Biochemistry 18: 351
• 2 BergströM K, Lahnborg G. 1975; The effect of a major surgery, low dosis of heparin and thromboembolism on plasma antithrombin. Comparison of immediate thrombin inhibiting capacity and the antithrombin III content. Thrombosis Research 6: 223
• 3 BlombåCK B, BlombåCK M. 1956; Purification of human and bovine fibrinogen. Arkiv Kemi 10: 415
• 4 BlombåCK M, BlombåCK B, Olsson P, Svendsen L. 1974; The assay of antithrombin using a synthetic chromogenic substrate for thrombin. Thrombosis Research 5: 621
• 5 Breen FA, Tullis JL. 1968; Ethanol gelation: A rapid screening test for intravascular coagulation. Annals of Internal Medicine 69: 1197
• 6 Bruhn HD, Heimburger N. 1976. Das kalt-unlösliche Globulin bei metastasierenden Tumoren und Leukämien. Thrombosis et Diathesis Haemorrhagica. Suppl. 62 83
• 7 Budzynski AZ, Marder VJ. 1975; Determination of human fibrinopeptide A by radioimmunoassay in purified systems and in the blood. Thrombosis et Diathesis Haemorrhagica 34: 709
• 8 Fletcher AP, Alkjaersig N, O’BRIEN J, Tulevski VG. 1970; Blood hypercoagulability and thrombosis. Transactions Association of American Physicians 83: 159
• 9 Furthmayr H, Timpl R. 1971; Characterization of collagen peptides by sodium dodecylsulfate-polyacrylamide electrophoresis. Analytical Biochemistry 41: 510
• 10 Godal HC, Abildgaard U. 1966; Gelation of soluble fibrin in plasma by ethanol. Scandinavian Journal of Haematology 3: 342
• 11 Godal HC, Abildgaard U, Kierulf P. 1971. Ethanol gelation and fibrin monomers in plasma. Scandinavian Journal of Haematology. Suppl. 13 189
• 12 Gollwitzer R, Timpl R, Becker U, Furthmayr H. 1972; Chemical and immunological properties of reduced and alkylated polypeptide chains of bovine fibrinogen. European Journal of Biochemistry 28: 497
• 13 Graeff H, Von Hugo R, Hafter R. 1973; In vivo formation of soluble fibrin monomer complexes in human plasma. Thrombosis Research 3: 465
• 14 Heene DL, Matthias FR. 1973; Adsorption of fibrinogen derivatives on insolubilized fibrinogen and fibrin monomer. Thrombosis Research 2: 137
• 15 Von HUGO R, Hafter R, Stemberger A, Graeff H. 1975; Soluble fibrin monomer complexes demonstrated by agarose gel filtration and by adsorption on insolubilized fibrinogen. Thrombosis et Diathesis Haemorrhagica 34: 216
• 16 Latallo ZS, Wegrzynowicz Z, Budzynski AZ, Kopec M. 1971; a Effect of protamine sulphate on the solubility of fibrinogen, its derivatives and other plasma proteins. Scandinavian Journal of Haematology Suppl 13: 151
• 17 Latallo ZS, Wegrzynowicz Z, Teisseyre E, Kopec M. 1971; b Simple and rapid evaluation of the intravascular coagulation and fibrinolytic states by application of protamine sulphate and reptilase R. Scandinavian Journal of Haematology Suppl 13: 387
• 18 Lipinski B, Worowski K. 1968; Detection of soluble fibrin monomer complexes in blood by means of protamine sulfate test. Thrombosis et Diathesis Haemorrhagica 20: 44
• 19 Mosesson MW, Umfleet RA. 1970; The cold-insoluble globulin of human plasma. I. Purification, primary characterization and relationship to fibrinogen and other cold-insoluble fraction components. Journal of Biological Chemistry 245: 5728
• 20 Nossel HL, Yudelman J, Canfield RE, Butler Jr VP, Spanondis K, Wilner GD, Quereshi GD. 1974; Measurement of fibrinopeptide A in human blood. Journal of Clinical Investigation 54: 43
• 21 Nossel HL, Butler Jr VP, Wilner GD, Canfield RE, Harfenist EL. 1976; Specificity of antisera to human fibrinopeptide A used in clinical fibrinopeptide A assays. Thrombosis and Haemostasis 35: 101
• 22 Ødgard OR, Lie M, Abildgaard U. 1975; Heparin cofactor activity measured with an amidolytic method. Thrombosis Research 6: 287
• 23 Stemberger A, HöRMANN H. 1975; Heterogeneity of clottable fibrinogen isolated from plasma by affinity chromatography. Zeitschrift für Physiologische Chemie 356: 341
• 24 Stemberger A, HöRMANN H. 1976; Affinity chromatography of immobilized fibrinogen and fibrin monomer, II. The behavior of cold-insoluble globulin. Zeitschrift für Physiologische Chemie 357: 1003
• 25 Svendsen N, BlombåCK B, BlombåCK M, Olsson P. 1972; Synthetic chromogenic substrates for determination of trypsin, thrombin and thrombin-like enzymes. Thrombosis Research 1: 267