Efficient production of recombinant human factor VIII by co-expression of the heavy and light chains

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Abstract
We have developed a high-level expression system for human blood coagulation factor VID (FVUI) consisting of a 90 kDa heavy (H-)chain and an 80 kDa light (L-)chain. Two expression plasmids were prepared, one expressing the H-chain and the other expressing the L-chain. These recombinant plasmids were designed to produce each chain linked to short additional ammo acid residues derived from the FVm precursor sequence. Furthermore, Kozak's translation initiation consensus sequence was introduced into the start codon for the H-chain. These modifications have dramatically increased the levels of expression of these chains. Chinese hamster ovary (CHO) cells co-transfected with these two recombinant plasmids were subjected to gene amplification and cloning. The final cell line, designated CTC-CF8, secretes 15 IU/day/106 cells of active FVm which is indistinguishable from plasma-derived FVm in its structure and biochemical properties. This system is suitable for large-scale production of pathogen-free recombinant human FVm which can be used for the treatment of haemophilia A patients.

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