US 10,590,405 B2 · Method for modifying human cell lines to produce factor VII

Abstract

A process for producing blood coagulation Factor VII in 3 human cell lines (HepG2, Sk-Hep, HKB-11) and to select the best recombinant protein producer is described. The murine line BHK-21 was used as control. The data allow to assert that the system used to modify cell lines was efficient, so that all the cells were satisfactorily modified, and produced the protein of interest of stable form. In addition, when comparing the murine line BHK-21 with the human cells (HepG2, Sk-Hep-1 and HKB-11), the latter showed to be able to produce rFVII more efficiently, which allows to conclude that human cell lines are a great alternative for the production of recombinant blood coagulation factors.

Claims14

Dependent claims

A process for modifying human cell lines to produce increased levels of blood coagulation Factor VII, comprising the steps of:

(1) providing virus particles containing a Factor FVII gene and a GFP gene as a reporter;
(2) transducting human cell lines selected from SK-Hep 1, HKB11 and HepG 2 with the viral particles to form human Factor FVII producing cells;
and
(3) culturing the Factor human FVII producing cells from step (2) in suspension using microcarriers, wherein in the process, expression of calumenin, VKORC1 and gamma-carboxylase genes, is not altered.

1 more element

Dependencies are inferred from the claim text.

Method for modifying human cell lines to produce factor VII

Life of this patent

Method for modifying human cell lines to produce factor VII

Life of this patent

Abstract

Claims14