Materials
Dried sea cucumbers Holothuria fuscopunctata were purchased from local markets in Zhanjiang, Guangdong Province, China. Amberlite FPA98Cl ion exchange resin was purchased from Rohm and Haas Company (USA). Polyacrylamide gels (Bio-Gel P2, P6 and P10) were from Bio-Rad (Bio-Rad Laboratories). Sephadex G-25 and G-10 were from GE Healthcare. Deuterium oxide (99.99%) was from Sigma Aldrich. LMWH (Enoxaparin, 0.4 mL 4000 AXaIU, Mw ~4.5 kDa) was from Sanofi-Aventis (France). Dermatan sulfate (DS, Mw 41.4 kDa) was from Sigma-Aldrich (USA). Oversulfated chondroitin sulfate (OSCS) was from National Institutes for the Control of Pharmaceutical and Biological Products (China). The activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) reagents, and human coagulation control plasma were all purchased from Teco Medical (Germany). Biophen f.VIII: C kit, Biophen antithrombin III (AT-III)-IIa kit, Biophen AT-III-Xa kit, Biophen VII kit, human AT-III, human factor VII (f.VII), human activated factor X (f.Xa), human thrombin (activated factor II, f.IIa), thrombin chromogenic substrate CS-01(38) and kallikrein chromogenic substrate CS-31(02), CS 11(65) were all from Hyphen BioMed (France). Recombinant coagulation f.VIII was from Bayer HealthCare LLC (USA). Human AT-III deficiency plasma was from Sekisui Diagnostics (USA). Human heparin cofactor II (HCII) deficient plasma was from Affinity Biologicals Inc. (USA). Human f.XIIa and activated factor VII (f.VIIa) was from Enzyme Research Laboratories (USA). Human f.XIa was from Assaypro (USA). Human f.IXa and human factor X (f.X) were from Haenatologic Technologies (USA). S-2366 was from Chromogenix (Italy). Pefchrome f.IXa and f.VIIa was from DSM (Netherlands). Hirudin was from Tian Enze Gene Technology (China), Amine-PEG 3 -Biotin was from MedChemExpress LLC (USA). SA biosensor was from Fortébio (USA). All other chemical reagents were of analytical grade and obtained commercially.