title: SK-HEP cells and lentiviral vector for production of human recombinant factor VIII.
abstract: Hemophilia A is caused by a deficiency in coagulation factor VIII. Recombinant factor VIII can be used as an alternative although it is unavailable for most patients. Here, we describe the production of a human recombinant B-domain-deleted FVIII (rBDDFVIII) by the human cell line SK-HEP-1, modified by a lentiviral vector rBDDFVIII was produced by recombinant SK-HEP cells (rSK-HEP) at 1.5-2.1 IU/10(6) in 24 h. The recombinant factor had increased in vitro stability when compared to commercial pdFVIII. The functionality of rBDDFVIII was shown by its biological activity and by tail-clip challenge in hemophilia A mice. The rSK-HEP cells grew in a scalable system and produced active rBDDFVIII, indicating that this platform production can be optimized to meet the commercial production scale needs.
authors.affiliation: Center for Cell Therapy and Regional Blood Center of Ribeirão Preto, University of São Paulo (USP), Ribeirão Preto, 14051-140, Brazil. nathaliagrosa@gmail.com
authors.forename: Nathalia Gonsales
authors.initials: NG
authors.lastname: da Rosa
authors.forename: Kamilla
authors.initials: K
authors.lastname: Swiech
authors.forename: Virgínia
authors.initials: V
authors.lastname: Picanço-Castro
authors.forename: Elisa Maria de Sousa
authors.initials: EM
authors.lastname: Russo-Carbolante
authors.forename: Mario Abreu
authors.initials: MA
authors.lastname: Soares Neto
authors.forename: Andrielle
authors.initials: A
authors.lastname: de Castilho-Fernandes
authors.forename: Vitor Marcel
authors.initials: VM
authors.lastname: Faça
authors.forename: Aparecida Maria
authors.initials: AM
authors.lastname: Fontes
authors.forename: Dimas Tadeu
authors.initials: DT
authors.lastname: Covas
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002460:Cell Line; D004195:Disease Models, Animal; D005169:Factor VIII / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D006467:Hemophilia A / Q000188:drug therapy; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D016019:Survival Analysis
chemical_list: C451034:B-domain-deleted factor VIII; D010446:Peptide Fragments; D011994:Recombinant Proteins; D005169:Factor VIII