Erythropoietin enhancer stimulates production of a recombinant protein by Chinese hamster ovary (CHO) cells under hypoxic condition

Abstract

Oxygen is a limiting nutrient in animal cell culture and its supply is still worthy of improvement for production of useful proteins with a high efficiency. From a different point of view, development of the system by which a high productivity can be maintained even under hypoxic condition as well as under normoxic condition may be important. A number of hypoxia-inducible genes have been found in eucaryotic cells and the induction in most cases, if not all, is due to hypoxic activation of the gene transcription. Transcription of erythropoietin gene is highly hypoxia-inducible and the induction is achieved by binding of a protein, which is widely distributed in animal cells, to a short DNA stretch (erythropoietin enhancer) in the 3' flanking region of erythropoietin gene. Using a hepatoma cell line (Hep3B) that produces the endogenous erythropoietin in an oxygen-dependent manner and Chinese hamster ovary cells that have been widely used for production of recombinant proteins, we show that, under hypoxic condition, the erythropoietin enhancer can activate not only the promoter of erythropoietin gene but also promoters of cytomegalovirus early genes and eucaryotic polypeptide chain elongation factor gene, both of which are very active in animal cells under normoxic condition.