Reactivity of Fibrinogen Derivatives with Antisera to Human Fibrin D-Dimer and Its γ-γ Chain Remnant

Summary

Highly purified D-dimer was obtained from plasmin digest of human cross-linked fibrin. After reduction of its disulfide bonds, the γ-γ chain remnant, containing cross-linking site, was then isolated by ion-exchange chromatography on CM-cellulose. Antisera obtained by immunizing rabbits with D-dimer and its γ-γ chain remnant contained a small population of antibodies which specifically reacted with D-dimer. Thus, a specific radioimmunoassay system allowing detection and quantitation of D-dimer in the presence of fibrinogen and monomeric fragment D was made possible.

Keywords

Fibrin D-dimer - Neodeterminants - Radioimmunoassay analysis

• References

• 1 Gaffney PJ, Brasher M. Subunit structure of the plasmin induced degradation products of cross-linked fibrin. Biochim Biophys Acta 1975; 295: 308-313
• 2 Kopeć M, Teisseyre E, Dudek-Wojciechowska G, Kloczewiak M, Pankiewich A, Latallo ZS. Studies on the “double” D fragment from stabilized bovine fibrin. Thromb Res 1973; 2: 283-291
• 3 Pizzo SV, Taylor Jr IM, Schwartz ML, Hill RL, McKee PA. Subunit structure of fragment D from fibrimogen and cross-linked fibrin. J Biol Chem 1973; 248: 4584-4590
• 4 Hudry-Clergeon G, Paturel L, Susdllon M. Identification d’un complexe (D-D). E dans les produits de dégradation de la fibrine bovine stabilisée par le Facteur XIII. Path Biol Suppl 1974; 22: 47-52
• 5 Gaffney PJ. Distinction between fibrinogen and fibrin degradation products in plasma. Chim Acta 1975; 65: 109-115
• 6 Lane DA, Preston PE, VanRoss ME, Kakkar VV. Characterization of serum fibrinogen and fibrin fragments produced during disseminated intravascular coagulation. Br J Haematol 1978; 40: 699-710
• 7 Graeff H, Hafter R, von Hugo R. Molecular aspects of defibrination in a case of amniotic fluid embolism. Thromb Haemostas 1977; 38: 724-727
• 8 Francis CW, Marder VJ, Martin SE. Detection of circulating crosslinked fibrin derivatives by a heat extraction SDS gradient gel electrophoresis technique. Blood 1979; 54: 1282-1295
• 9 Budzyński AZ, Marder VJ, Parker ME, Shames P, Brizuela BS, Olexa SA. Antigenic markers of fragment D-D, a unique plasmin derivative of human cross-linked fibrin. Blood 1979; 54: 794-804
• 10 Lee-Own V, Gordon YB, Chord T. The detection of neoantigenic sites on the D-dimer peptide isolated from plasmin digested crosslinked fibrin. Thromb Res 1979; 14: 77-84
• 11 Doolittle RF, Schubert D, Schwartz SA. Purification of human fibrinogen. Arch Biochem Biophys 1967; 118: 456-467
• 12 Deutsch DG, Mertz ET. Isolation of human plasminogen. Science 1970; 170: 1095-1096
• 13 Olexa SA, Budzyński AZ, Marder VJ. Modification of high molecular weight plasmic degradation products of human cross-linked fibrin. Biochim Biophys Acta 1979; 576: 39-50
• 14 Heene DL, Mathias FR. Adsorption of fibrinogen derivatives on insolubilized fibrinogen and fibrin monomers. Thromb Res 1973; 2: 137-154
• 15 Nussenzweig V, Seligmann M, Pelmont J, Graber P. Les produits de dégradation du fibrinogène humain par la plasmine. I Séparation et proprietés physicochimiques. Ann Inst Pasteur 1961; 100: 377-389
• 16 Gardlund B, Kowalska-Loth B, Grondahl NJ, Blombäck B. Plasmic degradation products of human fibrinogen. I. Isolation and characterization of fragments E and D and their relation to disulphide knots. Thromb Res 1972; 1: 371-388
• 17 Okude M, Iwanaga S. Carboxyl-terminal residues of mammalian fibrinogen and fibrin. Biochim Biophys Acta 1971; 251: 185-196
• 18 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Arkiv Kemi 1956; 10: 415-443
• 19 Marder VJ, Shulman NR, Caroli WR. High molecular weight derivatives of human fibrinogen produced by plasmin. I. Physicochemical and immunological characterization. J Biol Chem 1969; 224: 3111-3119
• 20 Gollwitzer R, Timpl R, Becker V, Furthmayr H. Chemical and immunological properties of reduced and alkylated polypeptide chains of bovine fibrinogen. Eur J Biochem 1972; 28: 497-506
• 21 Itzhaki RF, Gill DM. A microbiuret method for estimating proteins. Anal Biochem 1969; 9: 401-410
• 22 McDonagh J, Messel H, McDonagh RF, Murano G, Blombäck B. Molecular weight analysis of fibrinogen and fibrin chains by an improved sodium dodecyl sulfate gel electrophoresis method. Biochim Biophys Acta 1972; 257: 135-142
• 23 McConahey PJ, Dixon FJ. A method for trace iodination of proteins for immunological studies. Int Arch Allergy Appi Immunol 1966; 29: 185-189
• 24 Plow E, Edgington TS. Immunobiology of fibrinogen. Emergence of neoantigenic expressions during physiologic cleavage in vitro and in vivo. J Clin Invest 1973; 52: 273-282
• 25 Purves LR, Lindsey GG, Franks JJ. Sites of D-domain interaction in fibrin derived D-dimer. Biochemistry 1980; 19: 4951-4058
• 26 Ferguson EW, Fretto LJ, McKee PA. A re-examination of the cleavage of fibrinogen and fibrin by plasmin. J Biol Chem 1975; 250: 7210-7218