A novel anti-tissue factor monoclonal antibody with anticoagulant potency derived from synthesized multiple antigenic peptide through blocking FX combination with TF.
Tissue factor (TF) has been implicated in the pathogenesis of various thrombotic disorders. Monoclonal antibodies (mAb) that specifically target TF may have potential as antithrombotic therapy. We designed a unique TF peptide (TFP) that was specific for the binding site to factor X (FX). This peptide was used to develop TF mAb that block the coagulation cascade by interfering with the combination of FX with the TF/FVIIa complex. Chemically synthesized TFP coupled to polylysine matrix was used as multiple antigenic peptide (TF-MAP) and this was used to immunize Balb/c mice for the preparation of hybridomas. One hybridoma cell line released an antibody, named TF4A12, which had high anticoagulant potency (by dilute prothrombin time assay). Western blotting showed that TF4A12 could bind TF-MAP and the soluble TF extracellular domain (sTF(1-219)). Results of FX activation assay and amidolytic activity assay showed that the anticoagulant ability of TF4A12 is due to blocking FX, but not FVII, binding to TF. Our study identified an efficient method of developing TF mAb that could block the coagulation cascade.
Peng
Zhuo-Chun
ZC
Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Fudan University, 200032, People's Republic of China.
Cai
Xu
X
Zhang
Yu-Gao
YG
Kong
De-Sheng
DS
Guo
Hong-Shen
HS
Liang
Wang
W
Tang
Qi-Qun
QQ
Song
Hou-Yan
HY
Ma
Duan
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000494:pharmacology* / Q000627:therapeutic use; D000925:Anticoagulants; D000941:Antigens; D005170:Factor X / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology*; D011485:Protein Binding / Q000187:drug effects; D013925:Thromboplastin / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000925:Anticoagulants; D000941:Antigens; D010455:Peptides; D005170:Factor X; D013925:Thromboplastin