当前位置： SCI文献检索 > THROMBOSIS RESEARCH期刊下所有文献 > Enhanced functional recombinant factor VII production by HEK 293 cells stably transfected with VKORC1 where the gamma-carboxylase inhibitor calumenin is stably suppressed by shRNA transfection.

Enhanced functional recombinant factor VII production by HEK 293 cells stably transfected with VKORC1 where the gamma-carboxylase inhibitor calumenin is stably suppressed by shRNA transfection.

Abstract：

INTRODUCTION:Recombinant members of the vitamin K-dependent protein family (factors IX and VII and protein C) have become important pharmaceuticals in treatment of bleeding disorders and sepsis. However, because the in vivo gamma-carboxylation system in stable cell lines used for transfection has a limited capacity of post translational gamma-carboxylation, the recovery of fully gamma-carboxylated and functional proteins is low. MATERIALS AND METHODS:In this work we have engineered recombinant factor VII producing HEK 293 cells to stably overexpress VKORC1, the reduced vitamin K gamma-carboxylase cofactor and in addition stably silenced the gamma-carboxylase inhibitory protein calumenin. RESULTS AND CONCLUSIONS:Stable cell lines transfected with only a factor VII cDNA had a 9% production of functional recombinant factor VII. On the other hand, these recombinant factor VII producing cells when engineered to overexpress VKORC1 and having calumenin stably suppressed more than 80% by shRNA expression, produced 68% functional factor VII. The technology presented should be applicable to all vertebrae members of the vitamin K-dependent protein family and should lower the production cost of the clinically used factors VII, IX and protein C.

journal_name

Thromb Res

journal_title

Thrombosis research

authors

Wajih N,Owen J,Wallin R

doi

10.1016/j.thromres.2007.11.002

subject

Has Abstract

pub_date

2008-01-01 00:00:00

pages

405-10

issue

3

eissn

0049-3848

issn

1879-2472

pii

S0049-3848(07)00443-4

journal_volume

122

pub_type

杂志文章

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