Recent advances in technology supporting biopharmaceutical production from mammalian cells

Abstract

The demand for production of glycoproteins from mammalian cell culture continues with an increased number of approvals as biopharmaceuticals for the treatment of unmet medical needs. This is particularly the case for humanized monoclonal antibodies which are the largest and fastest growing class of therapeutic pharmaceuticals. This demand has fostered efforts to improve the efficiency of production as well as to address the quality of the final product. Chinese hamster ovary cells are the predominant hosts for stable transfection and high efficiency production on a large scale. Specific productivity of recombinant glycoproteins from these cells can be expected to be above 50 pg/cell/day giving rise to culture systems with titers of around 5 g/L if appropriate fed-batch systems are employed. Cell engineering can delay the onset of programmed cell death to ensure prolonged maintenance of productive viable cells. The clinical efficacy and quality of the final product can be improved by strategic metabolic engineering. The best example of this is the targeted production of afucosylated antibodies with enhanced antibody-dependent cell cytotoxicity, an important function for use in cancer therapies. The development of culture media from non-animal sources continues and is important to ensure products of consistent quality and without the potential danger of contamination. Process efficiencies may also be improved by employing disposable bioreactors with the associated minimization of downtime. Finally, advances in downstream processing are needed to handle the increased supply of product from the bioreactor but maintaining the high purity demanded of these biopharmaceuticals.

Tailoring recombinant protein quality by rational media design

“…They encompass several rodent-derived cell lines Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK) cells, and mouse myeloma cells, including NS0 and SP2/0, as well as human cell lines (HEK293, HT-1080) 6 . Chinese hamster ovary (CHO) cells are the most used mammalian cell line 7 , accounting for the production of more than 60% of mammalian cell culture derived currently approved biologics 6 . Very well characterized, CHO are likely to stay the expression system of choice as they have been successfully used to produce a variety of clinical biopharmaceuticals 7 .…”

“…Chinese hamster ovary (CHO) cells are the most used mammalian cell line 7 , accounting for the production of more than 60% of mammalian cell culture derived currently approved biologics 6 . Very well characterized, CHO are likely to stay the expression system of choice as they have been successfully used to produce a variety of clinical biopharmaceuticals 7 . A record of 12 novel biologics license applications (BLAs) were approved only in 2014 5 While the basic concepts of cell culture have remained unchanged since the mid-1980s 8 , improvements in the media compositions, including complex feeds opened the way for a dramatic yield increase 9 .…”

“…Both selection and engineering of the host cell line, as well as the culture conditions including media composition, shape the protein functionality 7,45 and effect undesired by-products, such as aggregates 46 and low-molecular-weight species (LMW) 47 . The choice of the host cell is decisive, since each of the frequently used cell lines (CHO, NS0 and SP2/0) has a cell-line specific glycosylation fingerprint.…”

“…Nowadays, more than 50% of the approved recombinant therapeutics are produced in mammalian cell lines, including Chinese hamster ovary cells, baby hamster kidney cells, and mouse myeloma cells, such as NS0 and SP2/0, as well as human cell lines 6 . Among them, CHO cells are the most frequently employed in the biotech industry, which represents more than 60% of the approved biologics that are produced in mammalian cell lines 6,7 . As a result of the high level of characterization of CHO and its successful use in the production of a variety of products over many years, it will likely continue, in the near future, to be the expression system of choice 7 .…”

“…Among them, CHO cells are the most frequently employed in the biotech industry, which represents more than 60% of the approved biologics that are produced in mammalian cell lines 6,7 . As a result of the high level of characterization of CHO and its successful use in the production of a variety of products over many years, it will likely continue, in the near future, to be the expression system of choice 7 . While the overall cell culture principles in industry have remained unchanged since the mid-1980s, improvements of media compositions, including complex feeds entailed dramatic yield increases, in particular in fed-batch processes 8,9 .…”