|
 |
The PER.C6® cell line development platform is designed to be user-friendly and robust, with rapid development timelines from transfection to stable cell line. All steps are performed in serum-free, chemically defined medium.
- Suspension-adapted host cells – no need to adapt to suspension culture during cell line development
- Hundreds of clones screened, using standard mammalian cell line development strategies
- Rapid development timelines – transfection to Research Cell Bank in 5 months
- High productivities, without amplification – cell lines with cell specific productivities of >50 pg/cell/day!
- PERCIVIA offers a full suite of cell line development support including vector construction, serum-free cell line generation up to Research Cell Bank, and cell line stability studies (>50 generations in culture, in the presence and absence of selective pressure). Research Cell Banks are tested for mycoplasma and sterility. A cGMP Master Cell Banking service is also available, through PERCIVIA’s partnership with DSM Biologics. In addition, full support and hands-on training on the use of PER.C6® cells for cell line development is available to all licensees.
[ back to top ]
Cell Culture: Batch and Fed-Batch Production Process
One of the key advantages of the PER.C6® technology is
the high performance of its platform production process. An ongoing
area of development at PERCIVIA is in the continuing improvement of this
baseline batch and fed-batch processes for material generation in support
of pre-clinical and clinical research.
Some of the key features of the platform batch and fed-batch production
processes include:
- High performance – PER.C6® cells may be cultured
to very high concentrations in batch and fed-batch cultures. Viable
cell concentration (Xv) > 12 X 106 cells/mL in batch and > 25
X 106 cells/mL in fed-batch are typically achieved (Figure 1). With
these high cell concentrations, integral viable cell (IVC) of over
100 billion cell days per liter (bcdl) in batch and over 300 bcdl in
fed-batch systems are routinely achieved. This translates to
productivities of ~ 2 g/L in batch and ~ 8 g/L in fed-batch with clones whose productivities average less than 30 picograms/cell/day
(pcd)
- Safety – all the media and supplements used in batch and fed-batch
processes are animal-derived component free and chemically defined.
- Easy implementation – the processes are designed to be easy
to implement in most mammalian cell culture labs and manufacturing
plants. The processes are simplified to require minimal operator
intervention and should be easily recognizable to scientists and operators
familiar with standard mammalian cell culture techniques and practices
 |
 |
| Typical Xv profiles of PER.C6® cells
in batch (red circle) and fed-batch (blue square) processes |
Typical product titer profiles of PER.C6® cells in batch
(red circle) and fed-batch (blue square) processes |
[ back to top ]
XD™ Process
The XD™ process takes advantage of the PER.C6® cell’s inherent
ability to withstand high shear and grow to very high concentrations. This
process uses Refine’s Alternating Tangential Flow (ATF) system
to perfuse fresh medium through the bioreactor and remove metabolic by-products
while retaining the cells. The result is a process in which PER.C6® cells
may be cultured to extreme densities and very high product concentration.
The features of the XD™ process include the following:
- Extreme cell concentrations – by continuously perfusing
in fresh nutrients and removing metabolic by-products, PER.C6® cells
are kept growing exponentially throughout the process. Typically
viable cell concentration of 100 – 150 X 106 cells/mL is normally
achieved in ~ 2 weeks
- Extreme productivity – Typical process IVC is ~ 500 bcdl in
two weeks. This means, with a cell line of typical productivity
(30 pcd), product concentration of10 - 15 g/L may be routinely achieved
- Consistency – continuous perfusion of fresh medium and removal
of metabolic byproducts means the cells are kept in a relatively constant
environment throughout the process. This results in more consistent
product quality profile and culture performance may be easily predicted
using a relatively straight-forward simulation model
 |
Typical Xv profiles of PER.C6® cells in XD™ processes compared to a typical fed-batch Xv profile. PER.C6® cells routinely reach > 120 X 106 cells/mL and often exceed 150 X 106 cells/mL approximately 2 weeks after inoculation in the XD™ process.
|
 |
| Typical product titer profile of PER.C6® cells in XD™ process compared to a typical fed-batch profile. Depending on the specific productivity of the clones used in the process, volumetric productivity of > 25 g/L of working volume (35 – 40 g/L in supernatant) is routinely achieved in this process approximately 2 weeks after inoculation. |
[ back to top ]
Protein Purification
Downstream process development is an integral part of the PER.C6® platform. Its focus is the use of economic and scalable technologies that enable the utilization of the full potential for this cell line and result high quality protein. PERCIVIA’s DSP philosophy is to creatively merge established technologies as well as breakthrough ones such as single use membrane chromatography techniques
- Design robust, cost effective, scalable platform processes for the manufacturing of Mabs and other recombinant molecules
- Accommodate high titer (> 5 g/L), and high cell density (> 30 million cells/mL) cultures
- Effective and scalable methods to harvest extreme cell density cultures (> 100 million cells/mL)
- Incorporates high capacity chromatography media (> 90 mg/L DBC for Mabs) and other breakthrough technologies
|
 |
Evolution of Downstream Process Unit of Operation-P’s
philosophy and direction. P’s Solution... Lower
Cost of Goods! |
[ back to top ]
|
|
