Efficient expression of functional human coagulation factor IX in stably-transfected Drosophila melanogaster S2 cells; comparison with the mammalian CHO system

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). Result Selection of an hFIX-expressing cell line derived from stably-transfected S...

Full description

Saved in:
Bibliographic Details
Published in:Biotechnology letters 2016-10, Vol.38 (10), p.1691-1698
Main Authors: Vatandoost, Jafar, Bos, Mettine H. A.
Format: Article
Language:English
Subjects:
Animals
Applied Microbiology
Barium
Biochemistry
Biomedical and Life Sciences
Biotechnology
Cell Line
Cellular biology
CHO Cells
Citrates
Coagulation
Cricetinae
Cricetulus
Drosophila
Drosophila melanogaster
Drosophila melanogaster - cytology
Drosophila melanogaster - genetics
Factor IX - genetics
Factor IX - metabolism
Glycosylation - drug effects
Hamsters
Hemophilia
Humans
Life Sciences
Mammals
Microbiology
Original Research Paper
Protein Engineering - methods
Protein expression
Recombinant
Secretions
Transfection
Tunicamycin - pharmacology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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). Result 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. Conclusion 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.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-016-2156-6