A novel function for selenium in biological system: selenite as a highly effective iron carrier for Chinese hamster ovary cell growth and monoclonal antibody production.
As the market for biopharmaceuticals especially monoclonal antibodies (MAbs) rapidly grows, their manufacturing methods are coming under increasing regulatory scrutiny, particularly due to concerns about the potential introduction of adventitious agents from animal-sourced components in the media used for their production in mammalian cell culture. Chinese hamster ovary (CHO) cells are by far the most commonly used production vehicles for these recombinant glycoproteins. In developing animal-component free media for CHO and other mammalian cell lines, the iron-transporter function of serum or human/bovine transferrin is usually replaced by certain organic or inorganic chelators capable of delivering iron for cell respiration and metabolism, but few of them are sufficiently effective. Selenium is a well-known essential trace element (TE) for cell growth and development, and its positive role in biological system includes detoxification of free radicals by activating glutathione peroxidase. In cell culture, selenium in the form of selenite can help cells to detoxify the medium thus protect them from oxidative damage. In this presentation, we describe the discovery and application of a novel function of selenite, that is, as a highly effective carrier to deliver iron for cell growth and function. In our in-house-developed animal protein-free (APF) medium for CHO cells, using an iron-selenite compound to replace the well-established tropolone delivery system for iron led to comparable or better cell growth and antibody production. A high cell density of >10 x 10(6) viable cells/mL and excellent antibody titer of approximately 3 g/L were achieved in 14-day fed-batch cultures in shake flasks, followed by successful scale-up to stirred bioreactors. The preparation of the commercially unavailable iron-selenite compound from respective ions, and its effectiveness in cell-culture performance, were dependent on reaction time, substrates, and other conditions.
Zhang
Jinyou
J
Bioprocess R&D, Merck & Co., Inc., Rahway, New Jersey 07065, USA. jinyou_zhang@alum.mit.edu
Robinson
David
D
Salmon
Peter
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D019149:Bioreactors; D001709:Biotechnology / Q000295:instrumentation* / Q000379:methods*; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods; D006224:Cricetinae; D005290:Ferric Compounds / Q000737:chemistry; D005973:Glutamine / Q000737:chemistry; D007501:Iron / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D012643:Selenium / Q000737:chemistry*; D018038:Sodium Selenite / Q000737:chemistry*; D014168:Transferrin / Q000737:chemistry; D014334:Tropolone / Q000737:chemistry
D000911:Antibodies, Monoclonal; D005290:Ferric Compounds; D011506:Proteins; D014168:Transferrin; D005973:Glutamine; C025314:ferric citrate; D014334:Tropolone; D007501:Iron; D012643:Selenium; D018038:Sodium Selenite