The monoclonal antibody that recognizes an epitope in the C-terminal region of the fibrinogen alpha-chain reacts with soluble fibrin and fibrin monomer generated by thrombin but not with those formed as plasmin degradation products

Thromb Res. 2007;121(3):377-85. doi: 10.1016/j.thromres.2007.05.008. Epub 2007 Jun 11.

Abstract

Introduction: The presence of soluble fibrin (SF) provides evidence of thrombin activation in the blood; therefore, SF is a useful marker for diagnosing blood coagulation diseases such as disseminated intravascular coagulation (DIC). The antibody that specifically detects SF could be a useful tool for diagnosing thrombotic diseases.

Materials and methods: By using an acid-solubilized desAA-FM (fibrin monomer) as an immunogen, we developed a monoclonal antibody, namely J2-23, which specifically reacts with SF and FM. We examined the specificity of J2-23 by ELISA and immunoblotting and confirmed the reactivity of J2-23 with SF and FM by gel filtration.

Results and conclusions: J2-23 specifically reacted with SF, but not with fibrinogen or plasmic fibrinogen-derived Fbg-X, Fbg-Y, Fbg-E, and D; thrombin-treated Fbn-X, Fbn-Y, and Fbn-E; and plasmic cross-linked fibrin (DD, XDP). The epitope recognized by J2-23 was located within the Aalpha 502-521 region on the C-terminal of the fibrinogen alpha-chain. The reactivity of J2-23 disappeared following the action of the fibrinolytic enzyme plasmin. Furthermore, J2-23 reacted not only with SF but also with FM in plasma from DIC patients. This indicated that J2-23 specifically detected coagulation without reflecting the plasmin action. We demonstrated the potential of J2-23 as a useful antibody for detecting SF for diagnosing blood coagulation.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity
  • Biomarkers / blood
  • Epitope Mapping
  • Epitopes / chemistry
  • Fibrin / chemistry
  • Fibrin / immunology*
  • Fibrin Fibrinogen Degradation Products / immunology
  • Fibrinogen / chemistry
  • Fibrinogen / immunology*
  • Fibrinolysin / metabolism
  • Humans
  • Hybridomas / immunology
  • Mice
  • Mice, Inbred BALB C
  • Solubility
  • Thrombin / metabolism
  • Thrombosis / blood
  • Thrombosis / diagnosis

Substances

  • Antibodies, Monoclonal
  • Biomarkers
  • Epitopes
  • Fibrin Fibrinogen Degradation Products
  • Fibrin
  • Fibrinogen
  • Thrombin
  • Fibrinolysin