title: Efficient expression of functional human coagulation factor IX in stably-transfected Drosophila melanogaster S2 cells; comparison with the mammalian CHO system.
abstract: OBJECTIVE
  To compare stably-transfected Drosophila melanogaster S2 and mammalian Chinese hamster ovary (CHO) cells for the expression and secretion efficiency of biologically-active human coagulation factor IX (hFIX).
  
  
  RESULTS
  Selection of an hFIX-expressing cell line derived from stably-transfected S2 cells was performed over 2 weeks, while the same procedure required 2 months for stably-transfected CHO cells. Furthermore, the selected S2 cell line was superior in producing of total hFIX protein (70 % increase), biologically-active hFIX (35 % increase), and specific hFIX activity (20 % increase) relative to the selected CHO cell line. Enrichment for functional, fully γ-carboxylated hFIX species via barium citrate adsorption demonstrated that up to 90 % of the hFIX expressed by S2 cells was γ-carboxylated versus 79 % of CHO-expressed hFIX. Inhibition of N-glycosylation by tunicamycin indicated that N-glycosylation of S2-expressed hFIX had occurred to a similar extent as in the CHO-produced hFIX.
  
  
  CONCLUSIONS
  The Drosophila S2 cell system is an attractive candidate for the efficient production of recombinant hFIX as it has the potential of significantly reducing the cell development time, while producing functional hFIX.
authors: Department of Biology, Hakim Sabzevari University, Sabzevar, Iran. j.vatan@hsu.ac.ir.
authors: Jafar
authors: J
authors: Vatandoost
authors: Department of Thrombosis and Hemostasis, Einthoven Laboratory for Vascular Medicine, Leiden University Medical Center, Leiden, The Netherlands.
authors: Mettine H A
authors: MH
authors: Bos
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics*; D005164:Factor IX / Q000235:genetics* / Q000378:metabolism*; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: D014415:Tunicamycin; D005164:Factor IX
keywords: CHO cells; Drosophila S2 cells; Hemophilia; Human factor IX; γ-Carboxylation