title: In vivo efficacy of human recombinant factor IX produced by the human hepatoma cell line HuH-7.
abstract: BACKGROUND
  Post-translational modifications of the CHO-cell-derived-recombinant human factor IX (FIX) currently used for the treatment of hemophilia B (HB) are different from plasma derived FIX. Our previous studies described a rFIX (HIX) having better profile of post-translational modifications than rFIX produced by CHO cells. The aim of the study consisted to verify the improved post-translational modifications effect of HIX on in vivo recovery.
  
  
  METHODS
  HIX has been produced in a bioreactor and then purified from supernatants. In vitro activation and activity were evaluated measured by thrombin generation tests (TGT) and compared to commercial molecules, Benefix(®) , Mononine(®) . The three molecules were then administrated (i.v.) to FIX-knockout mice and two minutes after injection, blood samples were collected and subjected to human FIX-specific-ELISA and TGT.
  
  
  RESULTS
  The clotting function of HIX, activation courses of HIX by FXIa and FVIIa-TF complex appear normal as did activation of Benefix(®) , Mononine(®) and TG constants of each FIX were equivalent. After injection to HB mice, circulating HIX did not present any significant difference in term of antigen value with Benefix(®) . Intriguingly, TGT were clearly exhibiting a better velocity for HIX than Benefix(®) and Mononine(®) . These data suggested that HIX may improve in vivo coagulant efficacy in comparison with the two commercial FIX injected at the same dose.
  
  
  CONCLUSIONS
  The study shows that HuH-7-derived-rFIX has better in vivo haemostatic activity in hemophilia B mice compared to the reference rFIX molecule despite similar in vivo recovery rates, suggesting that HuH-7 cells could represent an effective cellular system for production of rFIX.
authors.affiliation: Unite Hemostase, Inflammation and Sepsis EA 4174, Faculte de Medecine Laennec, Universite Lyon 1-Hospices Civils de Lyon, Lyon, France.
authors.forename: N
authors.initials: N
authors.lastname: Enjolras
authors.affiliation: Unite Hemostase, Inflammation and Sepsis EA 4174, Faculte de Medecine Laennec, Universite Lyon 1-Hospices Civils de Lyon, Lyon, France.
authors.forename: E
authors.initials: E
authors.lastname: Perot
authors.affiliation: Unite Hemostase, Inflammation and Sepsis EA 4174, Faculte de Medecine Laennec, Universite Lyon 1-Hospices Civils de Lyon, Lyon, France.
authors.forename: S
authors.initials: S
authors.lastname: Le Quellec
authors.affiliation: Unite Hemostase, Inflammation and Sepsis EA 4174, Faculte de Medecine Laennec, Universite Lyon 1-Hospices Civils de Lyon, Lyon, France.
authors.forename: A
authors.initials: A
authors.lastname: Indalecio
authors.affiliation: Unite Hemostase, Inflammation and Sepsis EA 4174, Faculte de Medecine Laennec, Universite Lyon 1-Hospices Civils de Lyon, Lyon, France.
authors.forename: J
authors.initials: J
authors.lastname: Girard
authors.affiliation: Unite Hemostase, Inflammation and Sepsis EA 4174, Faculte de Medecine Laennec, Universite Lyon 1-Hospices Civils de Lyon, Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Negrier
authors.affiliation: Unite Hemostase, Inflammation and Sepsis EA 4174, Faculte de Medecine Laennec, Universite Lyon 1-Hospices Civils de Lyon, Lyon, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Dargaud
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D003029:Coagulants / Q000097:blood / Q000627:therapeutic use; D004797:Enzyme-Linked Immunosorbent Assay; D005164:Factor IX / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000627:therapeutic use; D006207:Half-Life; D002836:Hemophilia B / Q000188:drug therapy / Q000662:veterinary; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D051379:Mice; D018345:Mice, Knockout; D011517:Prothrombin Time; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000097:blood / Q000627:therapeutic use
chemical_list: D003029:Coagulants; D011994:Recombinant Proteins; C093388:Mononine; D005164:Factor IX
keywords: HuH-7 cells; animal models; coagulation factor IX; haemophilia B; in vivo recovery; thrombin generation test