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

Abstract:

: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 gamma-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 gamma 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.

journal_name

Thromb Res

journal_title

Thrombosis research

authors

Lugovskoy EV,Gritsenko PG,Kolesnikova IN,Zolotarova EN,Chernishov VI,Nieuwenhuizen W,Komisarenko SV

doi

10.1016/j.thromres.2004.03.005

subject

Has Abstract

pub_date

2004-01-01 00:00:00

pages

251-9

issue

3-4

eissn

0049-3848

issn

1879-2472

pii

S0049384804001513

journal_volume

113

pub_type

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