title: 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.
abstract: BACKGROUND
  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.
  
  
  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.
  
  
  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.
authors.affiliation: Diagnostic Research Laboratories, Daiichi Pure Chemicals Co., Ltd., 3-3-1 Koyodai, Ryugasaki, Ibaraki 301-0852, Japan. suzuki-a@daiichichem.co.jp
authors.forename: Akiko
authors.initials: A
authors.lastname: Suzuki
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ebinuma
authors.forename: Masanao
authors.initials: M
authors.lastname: Matsuo
authors.forename: Osamu
authors.initials: O
authors.lastname: Miyazaki
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Yago
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015415:Biomarkers / Q000097:blood; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry; D005337:Fibrin / Q000737:chemistry / Q000276:immunology*; D005338:Fibrin Fibrinogen Degradation Products / Q000276:immunology; D005340:Fibrinogen / Q000737:chemistry / Q000276:immunology*; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D012995:Solubility; D013917:Thrombin / Q000378:metabolism; D013927:Thrombosis / Q000097:blood / Q000175:diagnosis
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D000939:Epitopes; D005338:Fibrin Fibrinogen Degradation Products; D005337:Fibrin; D005340:Fibrinogen; D013917:Thrombin; D005341:Fibrinolysin