OBJECTIVETo get stable cell line expressing B domain-deleted human FVIII (BDDhFVIII) by constructing the eukaryotic expression plasmid.

METHODSEukaryotic expression plasmid containing BDDhFVIII was constructed and transfected into HepG2 cells via electroporation. The expression and purification of the target protein was detected by Western blot.

RESULTSResults of enzyme digestion and sequence analysis demonstrated that the gene of BDDhFVIII was correctly inserted into the eukaryotic expression vector pcDNA4/v5-his. Western blot confirmed the successful expression of BDDhFVIII at the protein levels in HepG2 cells.

CONCLUSIONThe constructed eukaryotic expression vector was able to generate high level expression of human FVIII in HepG2 cells, thus could construct human blood coagulation FVIII stable cell line successfully.