title: [Development of a monoclonal antibody to factor VIII C2 domain and its functional study].
abstract: OBJECTIVE
  To develop a monoclonal antibody (mAb) directed to FVIII C2 domain and investigate its effect on FVIII activity.
  
  
  METHODS
  FVIII C2 protein was expressed in E. coli and purified. A murine antihuman FVIII C2 domain mAb SZ-132 was developed by standard hybridoma technology and characterized. In coagulation assays, different concentrations of SZ-132 were incubated with freshly collected pooled human plasma and the residual activity of FVIII and activated partial thromboplastin time (APTT) were determined. The effects of SZ-132 on rhFVIII binding to purified human vWF, phosphatidylserine (PS) and platelets were assessed by enzyme linked immunosorbent assays (ELISA).
  
  
  RESULTS
  SZ-132 could inhibit FVIII procoagulant activity in a dose-dependent manner within the concentrations of 0-25 microg/ml and the FVIII activity was completely inhibited on above 25 microg/ml. It could also prevent rhFVIII from binding to vWF, PS and platelets.
  
  
  CONCLUSIONS
  SZ-132 is a neutralizing mAb against FVIII C2 domain and can inhibit FVIII procoagulant activity by preventing FVIII from binding to vWF and PS.
authors.affiliation: First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis, Ministry of Health, Suzhou 215006, China.
authors.forename: Zhen-Yu
authors.initials: ZY
authors.lastname: Li
authors.forename: Yi-Ming
authors.initials: YM
authors.lastname: Zhao
authors.forename: Ning-Zheng
authors.initials: NZ
authors.lastname: Dong
authors.forename: Fei
authors.initials: F
authors.lastname: Shen
authors.forename: Chang-Geng
authors.initials: CG
authors.lastname: Ruan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000096:biosynthesis / Q000276:immunology; D005169:Factor VIII / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D005169:Factor VIII