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STRATEGIES FOR UPSTREAM INTENSIFICATION

Source: MilliporeSigma

By Habib Horry, Ph.D., Associate Director Marketing – Upstream Process
Integration, BioContinuum™ Platform and Melanie Brandl, Ph.D., Global Product
Manager Proprietary Media

Figure 1. The industry expects to significantly reduce manufacturing and capital
costs with process intensification (Adapted from BPOG Technology Roadmap, 2017).

The Facility of the Future represents a transformation in how biologics are
developed and manufactured. Upstream intensification is one strategy that will
enable this revolution by significantly driving down costs through an increase
in volumetric productivity. (Figure 1).

In addition to increasing productivity, upstream process intensification has the
potential to accelerate protein production and increase protein quality.
Perfusionbased strategies are increasingly leveraged for this intensification as
they deliver speed, productivity, flexibility and reliability. These strategies
can be applied to cell line development, seed train, production and cell culture
media formulation (Figure 2).



Figure 2. Strategies for upstream process intensification can be leveraged for
cell line development, the seed  train,production and cell culture media
formulation.

Streamlining Seed Trains with High Cell Density Cryopreservation (HCDC)

Thawing a single vial of cells to initiate cell expansion for a GMP
manufacturing batch is time-consuming and requires open cell culture operations.
Use of HCDC method, which feeds into the first seed train bioreactor, can
streamline the overall process by up to 10 days2 (Figure 3).



Figure 3. HCDC enables one seed train expansion step at a central site in
support of several production campaigns.

As the frozen volume is relatively high, single-use bags are more appropriate
than vials, allowing HCDC to be used in closed processing, reducing
contamination risks.

Significantly Improving Process Economics with a Perfused Seed Train

Perfusion-based processes, especially in the seed train, can reduce the costs of
manufacturing and increase product throughput, all while maintaining the
production bioreactor in a more simple-to-operate fed-batch mode.1

In addition, perfused seed trains allow for high seeding of the production
bioreactor, which can potentially enable an increase in titer, especially when
incorporating a robust media platform (Figure 4 and 5).



Figure 4. Upstream intensification approaches such as perfusion and HCDC can
improve process economics.

Achieving High Cell Densities and Productivity with Specially Designed Media
Platform

Specially designed cell culture media for perfused seed train cell expansion
allow for spiking the production bioreactor with higher cell densities.
Additionally, use of specially designed media platform across the workflow, from
cell bank to production, also creates a consistent environment for cells. This
minimizes the time needed for cell adaptation during transfers from the cryobank
to expansion and production (Figure 5).

Each formulation in the media platform has a specific role and must be
compatible with each other in order to minimize adaptation when transferring
cells from one step to the next:

 * Cryomedium must support freezing and thawing of cells without cell damage.
 * Expansion medium must have a high nutrient concentration to achieve low cell
   specific perfusion rates (CSPRs) at high cell densities, enabling greater
   seed train process efficiencies.
 * Perfusion medium must facilitate high productivity at low perfusion rates.



Figure 5. Media designed for upstream intensification support the overall
upstream process.

Getting the Most out of Perfusion Culture with Cell Retention Devices

An essential component for reducing the risk and complexity of perfusion culture
is a cell-retention device that allows for high cell density and a wide range of
media exchange rates, with minimal fouling and reproducible performance (Figure
6). Our cell retention device includes a flat-sheet membrane-based technology,
optimized to ensure performance, scalability and eliminate any potential
negative impact on cell health. Use of this technology results in greater
cell-mass production in the N-1 bioreactor allowing your production bioreactor
to be seeded at a higher cell density. This higher cell inoculation density
eliminates any lag phase resulting in a shorter time required to reach peak cell
density and achieving higher titers3.



Figure 6. A cell retention device is critical to derive the greatest benefit
from perfusion approaches.

Webinars
1. A Cost analysis and evaluation of perfused seed train scenarios through
process modeling. Utilization of process modeling to quantify the benefits of
perfused seed trains, and to determine under which scenarios these benefits
arise.
2. Media and process development for seed train intensification. Demonstration
of how combining media, specially designed for seed train, production and
harvest intensification, can increase the cell specific productivity (Qp) in the
final production stage.
3. Novel Perfusion Filter and Controller for N-1 Application .A study comparing
a control process to the use of tangential flow filtration in the seed train and
evaluating various process parameters.

EMDMillipore.com/webinar





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MORE FROM MILLIPORESIGMA

 * UPSTREAM INTENSIFICATION – ENABLING PERFUSION PROCESSES WITH CELL RETENTION
   TECHNOLOGIES
   
   In this white paper, we explore various strategies for the intensification of
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   viability of each solution.

 * HIGH CELL DENSITY CRYOPRESERVATION: A VALUABLE APPROACH TO SEED TRAIN
   INTENSIFICATION
   
   HCDC, an alternative to the traditional method for seed train expansion, can
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 * PREPARE YOUR CHO CELLS FOR HIGHER PRODUCTIVITY BY OPTIMIZING PERFUSED SEED
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 * PREPARING CHO CELLS FOR HIGHER PRODUCTIVITY BY OPTIMIZING A PERFUSED SEED
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   This app note describes how N-1 perfusion using an expansion medium supports
   inoculation of a fed-batch, production bioreactor at a much higher cell
   density resulting in increased product titers.

 * A COST ANALYSIS AND EVALUATION OF PERFUSED SEED TRAIN SCENARIOS THROUGH
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   A description of process modeling and comparison of process economics of a
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   in conventional and high-seed fed-batch...

 * CELLICON™ PERFUSION FILTER AND CONTROLLER DATA SHEET

 * SEED TRAIN INTENSIFICATION USING HIGH CELL DENSITY CRYOPRESERVATION AND
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 * EVALUATION OF NOVEL CELL RETENTION TECHNOLOGY AND CELL CULTURE MEDIA FOR N-1
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 * A PLATFORM STANDARD FOR VIRAL VECTOR MANUFACTURING AND COMMERCIALIZATION

 * INTENSIFIED BUFFER MANAGEMENT AS A STRATEGY FOR INCREASED MANUFACTURING
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 * BIOPROCESSING AND SCALEUP OF HEK293 CELLS FOR AAV PRODUCTION

 * NEXT-GENERATION PURIFICATION OF MONOCLONAL ANTIBODIES

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