title: Efficient stabilization of recombinant human coagulation factor VIII in the milk of transgenic mice using hFVIII and vWF co-expression vector transduction.
abstract: OBJECTIVE
  To investigate the reasons for the instability of human coagulation factor FVIII (hFVIII) in milk which is an intractable obstacle during the hFVIII production by a transgenic mammary gland bioreactor.
  
  
  RESULTS
  We constructed P1A3-hFVIIIBDD and P1A3-hFVIIIBDD-IRES-vWF co-expression cassettes for generating transgenic mice. P1A3-hFVIII/CMV-vWF double heterozygotes were also prepared by mating P1A3-hFVIIIBDD with CMV-vWF mice. hFVIII bioactivity in milk was determined under different storage conditions. The half-life (in vitro) of hFVIII bioactivity in P1A3-hFVIIIBDD-IRES-vWF mice was significantly longer than P1A3-hFVIIIBDD mice [77 ± 4.9 vs. 44 ± 2.6 h at 4 °C, 32.5 ± 5 vs. 19.7 ± 0.6 h at room temperature and 7.4 ± 1.4 vs. 3.4 ± 0.6 at 37 °C, respectively (P < 0.05)]. The half-life (in vitro) of hFVIII bioactivity in milk of double heterozygotes was similar to P1A3-hFVIIIBDD-IRES-vWF ones, demonstrating that the vWF transgene expression in hFVIII transgenic mice can efficiently improve the stabilization of hFVIII bioactivity in milk.
  
  
  CONCLUSIONS
  We provide a new approach of P1A3-hFVIIIBDD-IRES-vWF co-expression to generate more stable hFVIII in transgenic milk with rapid and low cost as well as valuable information for producing pharmaceutical proteins by transgenic mammary gland bioreactor.
authors.affiliation: Shanghai Institute of Medical Genetics, Children's Hospital of Shanghai, Shanghai Jiao Tong University School of Medicine, Shanghai, 200040, China.
authors.forename: Xiaoye
authors.initials: X
authors.lastname: Ren
authors.forename: Xiuli
authors.initials: X
authors.lastname: Gong
authors.forename: Qin
authors.initials: Q
authors.lastname: Cai
authors.forename: Xinbing
authors.initials: X
authors.lastname: Guo
authors.forename: Miao
authors.initials: M
authors.lastname: Xu
authors.forename: Zhaorui
authors.initials: Z
authors.lastname: Ren
authors.forename: Yitao
authors.initials: Y
authors.lastname: Zeng
mesh_terms: D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics*; D005169:Factor VIII / Q000032:analysis* / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006579:Heterozygote; D006801:Humans; D008822:Mice, Transgenic; D008892:Milk / Q000737:chemistry*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics; D013696:Temperature; D014161:Transduction, Genetic; D014841:von Willebrand Factor / Q000032:analysis* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D014841:von Willebrand Factor; D005169:Factor VIII