본원의 개시 내용은 재조합 단백질, 특히 rFVIII, B-도메인 제거된 rFVIII, rFIX 또는 rFVII/rFVIIa와 같은 응고 단백질의 세포발현을 개선시키기 위한, 세포 관류 배양과 함께 막-기반의 외부 세포 체류 시스템을 사용하는 새로운 용도를 제공한다. 이 시스템을 고 밀도의 세포에 사용하는 경우, 체류 시스템의 운영 중에 발생되는 기계적인 힘, 예를 들어 섬유를 통한 펌핑으로 발생되는 세포 순환으로 인해 보다 균질한 세포 배양이 달성된다.
The invention disclosed herein provides a novel use of an external membrane-based cell retention system in conjunction with perfusion cell culture for improved cell expression of recombinant proteins, particularly coagulation proteins such as rFVIII, B- Domain Deleted rFVIII, rFIX or rFVII/rFVIIa. The use of such a system at high cell density results in a more homogeneous cell culture due to mechanical forces induced during the operation of the retention system, such as the cell circulation induced by pumping through the fibers, in addition, the retention of some cellular and medium components allows an optimal environment to the cells. The use of the method provided results in improvements in cellular productivity and in volumetric productivity. The membrane based cell retention system is disposable and when connected to a disposable bioreactor, it allows the whole perfusion system to be disposable when the medium and the harvest are stored in disposable bags. The disposable perfusion bioreactor system can be operated as a closed system and does not require cleaning, steaming or the use of a hard piped facility.