Novel small scale TFF cell retention device for perfusion cell culture systems
© Martin et al. 2015
Published: 14 December 2015
Perfusion processes have traditionally been used for the generation of unstable proteins in cell culture systems. The use of perfusion for production of stable proteins has been limited by low product concentration, media costs, and system complexity. With the advent of new single-use technology and high producing cell lines, perfusion processes are gaining increased attention from industry. Also, the enhanced productivity of perfusion bioreactor compared to fedbatch enables use of smaller single-use systems, both for clinical scale production as well as potentially for manufacturing scale. These new perfusion processes cannot only be used as a production platform but also for process intensification of fed-batch processes. Perfusion can be used to dramatically increase the cell density of the N-1 bioreactor to accelerate the production process and achieve gains in efficiency.
The critical components of a perfusion system are the bioreactor controller, cell culture vessel, and cell retention device. The cell retention device or system is often used to define the type of perfusion system with gravity-based and filtration devices being the most common.
This study will show the application of a small-scale tangential flow microfiltration device(prototype small scale Prostak™) in a perfusion test platform. This prototype device was derived from EMD Millipore Prostak™microfiltration family of products typically used in the primary clarification of cell culture.
Materials and Methods
Perfusion Process Details.
Materials and Equipment
Vessel and Controller
3L Applikon® Glass Bioreactor
Single-use recirculation Levitronix™ Pump, Single-use Mobius® 20L bags (Harvest and Medium)
Cell Growth Monitoring - two methods
Vi-CELL® (Trypan blue exclusion method)
Biomass by capacitance (iBiomass™ - Fogale™)
Vessel Temp (C)
pH Set Point
7.0 ( ± 0.2)
Dissolved Oxygen (DO)
Air-Oxygen / Open Pipe
250 rpm (30W/m3)
Working Volume (L)
Daily Cell Bleed
Manually (w/ peristaltic pump)
We have demonstrated that a new small scale TFF device (ssProstakTM) can be used in perfusion applications. Currently bioreactor perfusion processes are used for number reasons including process intensification and continuous upstream. The data suggest the utility of ProstakTM in accomplishing these process goals. The performance metrics measured in this study are similar to those obtained with other perfusion processes using hollow fiber TFF cell retention devices, either normal flow or alternating.
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.