Biomedical Engineering Reference
In-Depth Information
5.
Biotechnol J
. 2011 Jan;6(1):56-65. A
single-use purification process
for the production of a
monoclonal antibody produced
in a PER.C6 human cell line.
Kuczewski M, Schirmer E, Lain
B, Zarbis-Papastoitsis G.
PERCIVIA LLC, 1 Hampshire St.,
Cambridge, MA.
mkuczewski@
Advances in single-use technologies can enable
greater speed, flexibility, and a smaller footprint for
multiproduct production facilities, such as at a
contract manufacturer. Recent efforts in the area of
cell line and media optimization have resulted in
bioreactor productivities that exceed 8 g/L in
fed-batch processes or 25 g/L in high-density cell
culture processes. In combination with the
development of single-use stirred-tank bioreactors
with larger working volumes, these intensified
upstream processes can now be fit into a single-use
manufacturing setting. Contrary to these upstream
advances, downstream single-use technologies have
been slower to follow, mostly limited by low
capacity, high cost, and poor scalability. In this study,
we describe a downstream process based solely on
single-use technologies that meets the challenges
posed by expression of a mAb (IgG(1)) in a high-
density suspension culture of PER.C6 cells. The cell
culture harvest was clarified by enhanced cell
settling (ECS) and depth filtration. Precipitation was
used for crude purification of the mAb. A high-
capacity chromatographic membrane was then used
in bind/elute mode, followed by two membranes in
flow-through (FT) mode for polishing. A proof of
concept of the entire disposable process was
completed for two different scales of the purification
train.
