Two monoclonal antibodies to D-dimer-specific inhibitors of fibrin polymerization

article






Lugovskoy, E.V.

Gritsenko, P.G.

Kolesnikova, I.N.

Zolotarova, E.N.

Chernishov, V.I.

Nieuwenhuizen, W.

Komisarenko, S.V.
D-dimer of human fibrin was used as antigen to obtain monoclonal antibodies (mAbs). We have obtained 16 hybridomas producing mAbs of different specificity. Only two of these mAbs inhibited fibrin polymerization. They are of the IgG-class. One mAb (II-4d) inhibited fibrin polymerization to 100% and another (II-3b) to 60% at a molar ratio mAb/fibrin=1.0. Fab-fragments of these mAbs inhibited fibrin polymerization completely at the same molar ratio. The epitopes for the mAbs studied are situated in the NH2-terminal part of the γ-chain in fibrin D-domain. Electron microscopy showed that fibrin was in monomeric form in the presence of these mAbs or their Fab-fragments. Thus, these mAbs stop the initial step of fibrin polymerization, i.e. protofibril formation. Only one site of protofibril formation is known now in COOH-terminal half of the D-domain γ chain named "a" site, which is complementary to the "A" site in the central E-domain of fibrin molecule. Our experiment with immobilized GPRP showed that the "a" site in fibrin D-fragment preserved its binding activity to GPRP when the D-fragment was complexed with mAbs-inhibitors of fibrin polymerization. Thus, these two mAbs inhibit fibrin polymerization not by blocking the sites "a" but either by blocking another (not "a") specific site in D-domain or by steric hindrance of highly organized fibrin polymerization process. © 2004 Elsevier Ltd. All rights reserved. Chemicals / CAS: fibrin, 9001-31-4; glycylprolylarginylproline, 67869-62-9; immunoglobulin G, 97794-27-9; Antibodies, Monoclonal; Biopolymers; Epitopes; Fibrin Fibrinogen Degradation Products; fibrin fragment D; Fibrin, 9001-31-4; Fibrinogen, 9001-32-5; glycyl-prolyl-arginyl-proline, 67869-62-9; Immunoglobulin Fab Fragments; Oligopeptides
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Biomedical ResearchFibrinMonoclonal antibodiesPolymerization inhibitorsPolymerization sitesD dimerepitopefibrinfibrin monomerglycylprolylarginylprolineimmunoglobulin F(ab) fragmentimmunoglobulin Gmonoclonal antibodypeptideprotein inhibitorunclassified drugalpha chainantibody combining siteantibody productionantibody specificityelectron microscopyfibrin formationfibrin polymerizationhybridomamoleculeprotein bindingprotein domainprotein immobilizationterminal sequenceAntibodies, MonoclonalAntibody SpecificityBiopolymersBlood CoagulationEnzyme-Linked Immunosorbent AssayEpitopesFibrinFibrin Fibrinogen Degradation ProductsFibrinogenHumansHybridomasImmunoglobulin Fab FragmentsOligopeptidesProtein Structure, Tertiary
TNO Identifier
237773
Repository link
ISSN
00493848
Source
Thrombosis Research, 113(3-4), pp. 251-259.
Pages
251-259
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