Biology Reference
In-Depth Information
Commercial production of recombinant
erythropoietins
Alice S. Chuck
1
, Rohini R. Deshpande
2
, Adrian R. Distler
3
, Shane A.
Sander
3
,and James E. Seely
4
1
Amgen Inc., ATO Process Development, One Amgen Center Drive, M/S 18S-1-A, Thousand Oaks,
California 91320, USA
2
Amgen Inc., Protein Science, One Amgen Center Drive, M/S 14-2-C, Thousand Oaks, California
91320, USA
3
Amgen Inc, ATO Process Development, One Amgen Center Drive, M/S 30W-3-A, Thousand Oaks,
California 91320, USA
4
Amgen Inc, ACO Process Development, M/S AC-24F, Longmont, Colorado 80503, USA
Introduction
For the production of recombinant products from genetically engineered cells, a
number of cell hosts may be used, but the ones generally used belong to one of
five categories: plant, bacterial, yeast, insect, or mammalian. Depending on the
type of product that is desired, the features of that product, the intended use of
the product, and the preferred method of manufacture, an appropriate host cell
can be selected. With the appropriate genetic engineering, the gene of interest
such as erythropoietin (EPO) can be produced by a host cell. In the case of EPO
production, the sequence of amino acids as well as the amount of glycosylation
must be correct to achieve the desired efficacy
in vivo
. This chapter describes
one method of EPO production using cells genetically engineered to secrete
recombinant human erythropoietin (rHuEPO). In this method of production,
mammalian cells, which are capable of producing glycosylated forms with the
desired efficacy in humans, are typically selected as hosts. The host mammalian
cells secrete the rHuEPO product into the medium environment in which they
are cultured, making the remainder of production a matter of separating the
rHuEPO product from the cells and other components in the cell culture broth.
This chapter summarizes the generation of recombinant rHuEPO-producing cell
lines, the production of rHuEPO, the separation of rHuEPO from components
of the cell culture broth, and the packaging of the final rHuEPO drug product.
Cell line development
The development of a cell line generates a consistent source of cells that is
capable of satisfying commercial demand and that adheres to regulatory guide-
