title: Affinity-purification of fibrinogenase with high proteolytic activity from Agkistrodon halys (Chinese) Venom.
abstract: To purify and characterize the fibrinogenase with high proteolytic activity from Agkistrodon halys (Chinese) Venom. Monoclonal antibodies against fibrinogenase were prepared and a novel affinity chromatography equipped with a monoclonal antibody against fibrinogenase was developed and applied for the purification of fibrinogenases. The purified fibrinogenase was identified by fibrinolytic activity assay, and antithrombosis activity assay. HPLC chromatography and SDS-PAGE analysis demonstrated the uniformity and purity of the purified fibrinogenase. In comparison with a conventional A-50 chromatography method, affinity-purified fibrinogenase showed higher activity (3631 U mg(-1) vs 501 U mg(-1)). In addition, the physiological activity of the fibrinogenase both in vitro and ex vivo showed the purified fibrinogenase can specifically degrade beta-, gamma-fibrinogen and has a high anti-thrombotic activity. In conclusion, the purified fibrinogenase by affinity column were shown to be homogeneous and showed a high and specific proteolytic activity against beta-chains of fibrinogen molecules and antithrombosis activity.
authors.affiliation: College of Food Science & Nutritional Engineering, China Agricultural University, Beijing, 100083, China.
authors.forename: Biao
authors.initials: B
authors.lastname: Ma
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Dan
authors.initials: D
authors.lastname: Wu
authors.forename: Jianping
authors.initials: J
authors.lastname: Jia
authors.forename: Wentao
authors.initials: W
authors.lastname: Xu
authors.forename: Yunbo
authors.initials: Y
authors.lastname: Luo
mesh_terms: D017836:Agkistrodon; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003636:DEAE-Cellulose; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005342:Fibrinolysis / Q000187:drug effects; D005343:Fibrinolytic Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D007120:Immunochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D014757:Viper Venoms / Q000201:enzymology*
chemical_list: D000911:Antibodies, Monoclonal; D005343:Fibrinolytic Agents; D014757:Viper Venoms; D003636:DEAE-Cellulose; D012697:Serine Endopeptidases; C412879:venombin B