Journal
BLOOD
Volume 108, Issue 12, Pages 3757-3760Publisher
AMER SOC HEMATOLOGY
DOI: 10.1182/blood-2006-02-004671
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Funding
- NHLBI NIH HHS [R01HL69331, R01 HL069331] Funding Source: Medline
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To improve production of functional fully gamma-carboxylated recombinant human clotting factor IX (r-hFIX), cell lines stably overexpressing r-hFIX have been engineered to also overexpress proteins of the gamma-carboxylation system. Here we demonstrate that siRNA silencing of calumenin, an inhibitor of the gamma- carboxylation system, enhances production of functional r-hFIX produced by engineered BHK21 cells. The production yield of functional r-hFIX was 80% in engineered cells where calumenin had been silenced 78%. We propose that this high-yield expression system can easily be adapted to overproduce functional forms of all members of the vitamin K-dependent protein family.
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