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March 2011 | Volume 9 | Supplement 1
Commercializing a New
Class of Biopharmaceuticals
MACS is a registered trademark of Miltenyi Biotec GmbH. Copyright 2011 Miltenyi Biotec GmbH. All rights reserved.
Enabling cells for therapy
macs-stemcells.com/ct
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t "EESFTTJOHBMMZPVSOFFETJOEFWFMPQNFOU
NBOVGBDUVSJOHBOEBOBMZTJT
Every cell therapy
process is unique.
....so is every
scale up solution.
NORTH AMERICA EUROPE ASIA PACIFIC
To nd out more call 866 969 3232, email info@invetech.com.au or visit www.invetech.com.au
Argos Therapeutics
An automated cellular and RNA processing system developed to manufacture
autologous immunotherapies to treat advanced kidney cancer (metastatic renal cell
carcinoma), B-cell chronic lymphocytic leukemia and HIV.
KBI Biopharma
kSep, a closed continuous centrifuge was developed to gently concentrate
capture and/or separate cells by providing a very low-shear and nourishing
environment.
Invetech is an innovative consulting and engineering service provider developing scale-up strategies, automated equipment and
consumables for the development and manufacture of biological products, including cell therapies. Combining a clients process knowledge
with Invetechs technology and automation expertise, systems are created to meet client specic requirements for quality, cost and scale-up.
Invetechs specialist capability across the full development journey, from idea to market, provides the condence needed to achieve a
successful commercial outcome.
Invetech. Successful cell therapy
commercialization solutions.
Developers of closed, automated solutions.
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2 BioProcess International MARCH 2011 SUPPLEMENT
VOLUME 9 SUPPLEMENT 1 MARCH 2011
From the Publisher and Editor . . . . . . . . . . . . . . . . . 3
PART 1: INTRODUCTION
Therapies of Tomorrow Require More Than
Factories from the Past . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
David James
PART 2: CLINICAL DEVELOPMENT, NEW PRODUCTS
Successful Commercialization Through Industry
Collaboration: An Interview with Robert Deans
of the International Society for Cellular Therapy.. . . . . . . .12
Maribel Rios
Opportunities in Regenerative Medicine:
The Global Industry and Market Trends . . . . . . . . . . . . . . . .14
R. Lee Buckler
Stem-CellBased Therapies: Whats in Development,
Implications for Bioprocessing . . . . . . . . . . . . . . . . . . . . . . . . .20
Robert Shaw
Technologies on the Cutting Edge . . . . . . . . . . . . . . . . . . . . . .26
Maribel Rios
PART 3: PROCESS AND PRODUCT DEVELOPMENT
Cell Therapy Bioprocessing: Integrating Process
and Product Development for the Next Generation
of Biotherapeutics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .30
Ralph Brandenberger, Scott Burger, Andrew Campbell,
Tim Fong, Erika Lapinskas, and Jon A. Rowley
Meeting the Challenges in Manufacturing
Autologous Cellular Therapies . . . . . . . . . . . . . . . . . . . . . . . . . .38
Tamara T. Monesmith
Industry Roundtable: Viewpoints on Processing,
Quality, and Regulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .42
Maribel Rios
PART 4: BUSINESS ASPECTS
Addressing Business Models, Reimbursement,
and Cost of Goods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .46
Dawn Driscoll
PART 5: INDUSTRY EDUCATION FOR THE FUTURE
Industry Educational Platforms Drive Commercialization
Objectives: An Interview with Tracie Lodie of the
International Society for Cellular Therapy . . . . . . . . . . . . . .50
Maribel Rios
Working Together for the Future: A Letter from the
Chair of the Alliance for Regenerative Medicine . . . . . . . .54
Gil Van Bokkelen
Cell Therapy Resources . . . . . . . . . . . . . . . . . . . . . . .56
ON THE COVER
Regenerative medicines
are in development for a
wide range of indications,
as seen in this design by
Yusef Ramelize and
Cheryl Scott using an image
from BioLife Solutions.
(WWW.BIOLIFESOLUTIONS.COM)
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MARCH 2011 BioProcess International 3 SUPPLEMENT
FROM THE PUBLISHER AND EDITOR
Building a Bridge to Commercial Success
The history of the biopharmaceutical industry is one of
continual invention and reinvention, of business models
that have adapted to weather uncertain product futures
and shifting economic fortunes. Some of us followed the
up-and-down (and often financially painful) progress of
monoclonal antibodies toward their eventual commercial
success a wealth of experience to draw from as other
classes of products make their way from laboratories and
onto the market.
The vast majority of regenerative medicines are still
produced at laboratory scale, with suppliers targeting the
current market: research hospitals and laboratories. These
therapies could transform our approach to healthcare
and have a profound effect on the biotechnology industry
as a whole. They have the potential to reap tremendous
commercial successes.
What has been lacking is a strategic bridge connecting
current lab-scale cell therapies to commercialization.
The core challenge faced by this market lies in securing
funding which in turn rests on the ability of
companies to present solid data to ensure and support
commercially viable processes. The days when business
plans were approved and money secured based on
exciting science and visionary stories (rather than
solid projections) are over. The economic downturn and
increased sophistication of potential investors have made
it harder for start-up companies to secure funding.
Funding is indeed available. But a company must
show a detailed business plan and corporate strategy
that express confidence in market opportunities, early
clinical successes, management transparency, and
disease amelioration as well as efficacy that compares
favorably with existing treatment modalities.
That is the purpose of this issue: To serve as an
educational bridge to help move cell therapies from
laboratory to commercial scales. BPI will therefore
continue to delve into the science, technology, and
business realities of regenerative medicine, a major part of
the biopharmaceutical industry that we support.
Hundreds of regenerative medicines are in
development, and a number of tissue therapies (regulated
for the most part as devices) are already doing well on
the market. The hoped-for tipping point may have been
reached in mid-2010 with the approval of Dendreons
Provenge cell therapy. That company continues to pave the
way by venturing into the reimbursement arena, helping to
educate regulators and policymakers, and clearing the path
for further innovative therapies. Companies poised on
the brink of commercial launch are leading by example to
show how business models for such therapies must diverge
from well-known protein/antibody models.
For regenerative medicine, cells themselves are the
product, and scale-out (rather than -up) is the task.
Commercial success for most such therapies cannot
rest on continuing to replicate the same manual cell
reprogramming processes over and over again. Few
if any of those processes are yet automated. One recent
example suggested that, to produce enough cells to treat
1,000 patients, you might need some 20 cleanrooms
with identical equipment and 100 staff. But to produce
enough of the same product to treat 10,000 patients, you
could need 200 cleanrooms with identical equipment
and 850 staff. Add to that the logistics involved in rapid
processing and shipment from and back to patients
of fragile and sometimes irreplaceable patient-specific
cells and an industry that initially looks familiar to
what we already know about bioprocessing becomes, at
commercial scale, something very very different.
So why is BPI publishing a cell therapy supplement?
First, we remind you of our tagline: Covering the whole
development process for the global biotechnology industry. BPI
focuses on the myriad processes of moving products from
discovery to production (not simply processing as a discrete
step). From the start weve looked at the entire scope
of the industry both CDER- and CBER-regulated
products waiting for a point at which the regenerative
industry is primed to go commercial.
Given the tremendous promise, the thought that
cell therapy companies might struggle to replicate the
hard-fought lessons of other industry segments is just
unacceptable. One essential task is to help educate
potential investors be they venture capitalists, private
investors, or pharmaceutical companies seeking to expand
their product portfolios through strategic partnering. Our
readers want to serve the needs of humanity; they are
passionate about the promise of eliminating the physical
and economic ravages of the most devastating diseases.
Without successful commercialization, however, ground-
breaking therapies cannot reach those who need them.
So we are delighted to introduce you to so many
dedicated people and the good work they are doing to
advance the future of healthcare. We are eager to help
this segment of the industry communicate with and
learn from the experiences of the others and thereby
help speed these life-saving products to the market.
And we are grateful to Bob Speziale and David James
of Invetech; Lee Buckler from the Cell Therapy Group;
and Jon Rowley of Lonza Walkersville the team who
encouraged us to develop this issue and advised us along
the way. We also thank Jane Arthurs, who introduced us
to the ISCT committees; the organizers of the Phaciliate
Cell and Gene Therapy Forum; and all the authors and
interviewees who assisted in putting this project together.
Brian Caine,
publisher
S. Anne
Montgomery,
editor in chief
4 BioProcess International MARCH 2011 SUPPLEMENT
CELL THERAPI ES INTRODUCTION
Therapies of Tomorrow Require
More Than Factories from the Past
by David James
L
ive cells are being
incorporated as active agents
and delivery vehicles for a
broad range of emerging
therapeutic strategies. Successful
commercialization of a cell therapy
requires more than proving its
safety and efficacy to regulators.
Ultimately a therapy must be
commercially viable, allowing
enough patients to be treated with
an adequate financial margin to
justify investment in it as a product.
Whether the cells used are
universal (allogeneic) or patient-
specific (autologous), it is unlikely
to be wholly one or the other that
will dominate (1).
Commercial viability of
conventional therapeutic products
rests on economies of scale.
Investment in plants, facilities, and
personnel can serve a significant
patient population with a defined
candidature profile for a given
therapy. Factories used to produce
high-volume products are based
around established, scalable processes
with batch-control and risk-
management strategies that focus on
monitoring product quality. A drug
manufacturer can change the
manufacturing process extensively
and analyze the finished product to
establish that it is the same (2).
By contrast, many emerging cell
therapies will serve relatively small
patient populations with
autologous therapies at the extreme,
requiring a discrete batch for each
patient. Furthermore, for biologics,
the product is the process. Because
the finished product cannot be fully
characterized in the laboratory,
manufacturers must ensure product
consistency, quality, and purity by
ensuring that the manufacturing
process remains substantially the
same over time (2). So attempts to
dedicate a conventional production
philosophy to producing small-batch
therapies involve huge investment in
cleanrooms and equipment that is
often dedicated to each single small
batch with robust segregation
policies and procedures (3). Facility
use is therefore poor, with extensive
change-over costs when equipment
is cleaned and prepared to switch to
the next product batch.
Processing technologies used
during cell therapy clinical trials are
typically based on manual laboratory
methods that are not optimized or
easily integrated for production.
Further, although many cell
therapies in development require
similar processes, in reality such
novel therapies often require equally
novel processing technologies.
Those processes also require
significant manual interaction by
highly trained, professional staff.
Bench-level cellular therapeutic
production in the preclinical stage
requires skilled and highly focused
personnel working for long hours in
cleanroom conditions.
If you watch a manual cellular
process, you will see many detailed
manual operations: transferring
f luids from vessel to vessel, taking
cell counts, controlled dilutions, and
so on (Photo 1). Even if those
cleanrooms are built and the staff
well trained, maintaining a level of
consistency and quality to satisfy
regulatory requirements and process
demands is challenging.
Consequently, many small-batch cell
therapies have manufacturing
processes that are not well suited to
a current good manufacturing
practice (CGMP) environment. The
cost of such therapies can be
prohibitive, and managing scale-up
to meet demand is a particularly
daunting proposition (Figure 1).
Autologous therapies appear to
hold the key to controlling
communicable disease transmission
and addressing the need for immune
suppression, both of which limit the
application of allogeneic cell
therapies. Despite those potential
Photo 1 WESTMEAD HOSPITAL WWW.SWAHS.HEALTH.NSW.
GOV.AU/WESTMEAD/INDEX.HTM
MARCH 2011 BioProcess International 5 SUPPLEMENT
benefits, high costs normally
associated with small-batch
manufacturing often compromise its
commercial viability and can be a
major disincentive to investors and
developers. Herein lies a conundrum
for many companies interested in
developing new cell therapies:
Traditional bioprocessing factories
are not appropriate for small-batch
processes. Consequently, many cell
therapies are developed using
technologies and procedures that are
inherently expensive and labor-
intensive and require a high degree
of skill, training, and management
to minimize deviations.
To develop alternative processes
that are scalable will typically require
significant investment over a
considerable period. To prevent
complications associated with
postapproval process changes, a new
process needs to be ready for use
before clinical trials are complete
well before it is certain that the
therapy will be approved for market.
Several cell therapy companies have
recognized the need to address this
issue. Accordingly some have invested
a proportion of funds allocated for
clinical trials to planning, developing,
and optimizing innovative,
customized solutions to their
manufacturing challenges.
THE ROLE OF QBD AND PAT
There is an obvious commercial
imperative for small-batch cell
therapies, but process changes cannot
be made without careful
consideration. Biological products can
be sensitive to very minor
manufacturing process changes. Even
relatively small process differences
can significantly affect the nature of
a finished product most
importantly, how it functions in a
patients body. When the process is
the product as would be the case
with industrialized production of
stem cells variability is the enemy
and must be reduced and controlled
as completely as possible (4).
Successful biological
manufacturing therefore addresses
scale-up, process comparability, and
process characterization. Because
cell therapy products cannot be fully
characterized in a laboratory,
manufacturers must ensure product
consistency, quality, and purity by
ensuring that their manufacturing
processes remain substantially the
same over time. So companies must
tightly control the source and nature
of their starting materials and
establish appropriate process
controls for each unique product
and/or manufacturing process.
Further complicating these
matters is the fact that during
research and early stage clinical
trials, most cell therapies and
associated process controls are
developed and produced for a
limited number of patients. They
use technologies that are readily
available rather than what will
ultimately be needed for
commercial-scale production.
Typically those are manual,
laboratory-scale technologies that
are not optimized for specific cells
or process needs.
Authors from Aastrom
Biosciences, Inc. (www.aastrom.com)
highlighted in 2008 the criticality of
batch failure for autologous therapies:
The risk that a process will fail to
yield a finished cell dose meeting
specifications must be extremely low,
whatever the reason. For PSCT,
manufacturing failure is not merely
an operational inefficiency issue;
Photo 2: Sterile connections
for a closed consumable
Photo 3: QC sample collection
for a closed consumable
Figure 1: Schematic representation of scale-up for a typical manual cell therapy process; as the
number of patients increases, so must the number of technicians to prepare their treatments
IND
10 patients
Phase 1
10 patients
Phase 2
20 patients
Phase 3
50 patients
Commercialization
100 patients 1,000 patients 10,000 patients
6 BioProcess International MARCH 2011 SUPPLEMENT
instead, it directly translates into a
failed patient treatment (5). By
adopting a quality by design (QbD)
approach to developing cell therapy
processes, we can identify and
address critical failure modes and
process parameters.
For example, one useful measure
of process risk is the number of sterile
connections that must be made
throughout cell therapy
manufacturing. In a typical manual
process, operators make multiple
sterile connections in a biological
safety cabinet (BSC) over several days
to add reagents, collect quality
control (QC) samples, and package a
final product. In one relatively simple
process we analyzed recently, we
counted 76 sterile connections, all of
which were performed manually.
Each connection carries an inherent
risk to the product. When patient
numbers are small, such risk is
theoretically manageable by skilled
operators using good standard
operating procedures (SOPs). But risk
to products is considerable in
commercial-scale production.
A natural consequence of the QbD
approach to redesigning such a process
would be to minimize the number of
manual sterile connections. In fact,
when this concept of minimizing sterile
connections is taken to the extreme, it
implies a process without any manual
sterile connections. It is therefore
hardly surprising that much innovation
in small-batch manufacturing for cell
therapies has revolved around using
presterilized closed consumables
(single-use technology).
Process analytical technology
(PAT) has been defined as a system
for designing, analyzing, and
controlling manufacturing through
timely measurements (that is, during
processing) of critical quality and
performance attributes of raw and
in-process materials and processes,
with the goal of ensuring final
product quality (7). This philosophy
for monitoring and controlling
product quality is independent of
batch size, and its implications can be
profound as those batch sizes reduce.
For example, consider QC for an
autologous therapy. Regulators are
likely to require that the same tests be
performed regardless of batch size.
Each QC sample must be a specific
volume to suit a given analytical
method. Due to economies of scale,
the volume of cells used for a large
batch and the subsequent cost of
performing QC tests are relatively
minor. However, for an autologous
therapy, such identical QC samples
are likely to represent a significant
proportion of the total cells available
and the total cost per patient.
Final-product release testing must
satisfy a difficult combination of
constraints. Such tests must be
rapid (for products with short
shelf-lives), relatively inexpensive
(because they must be performed
for every dose), limited in
necessary sample volume (to
prevent excessive loss of final
product), and able to test a
complex product composition. (5)
So for small batches, it is even
more important that we invest efforts
in characterizing processes so that we
understand precisely which
parameters need to be monitored,
controlled, and tested including
operating limits within which a
predictable outcome can be assured.
If we combine the need to take
samples for PAT and QC with the
objective to use closed consumable
processing, then we have a specific
small-batch requirement for which a
solution must be found. How do we
efficiently collect samples from the
right place, at the right time, without
affecting product sterility (Photo 2)?
CLOSED SYSTEMS
Segregation is the comprehensive,
verifiable isolation of an internal
process from external contamination
at all points in a processing system.
Figure 2: Typical elements of a closed and automated cell processing system
Disposable processing set
Intermediate patient-specifc
product
Reagents and bufers
Automated processing
platform in a Class C area
In-process quality control
sample processing
Patient-specifc product
output to incubation,
freezing, and so on
Batch record
Figure 3: Typical facility layout for closed and automated cell therapy manufacturing
Reagents Store
Consumables
Store
Reagents
Prep Suite
Incubation
Patient-Specifc
Materials Prep
Process Recovery
Suite
Processing
Room
Quality Control
Freezing
Waste
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8 BioProcess International MARCH 2011 SUPPLEMENT
All process equipment and materials
must be cleaned and sanitized or
sterilized between batches to prevent
contamination. Cleaning and
sanitization/sterilization processes
also need to be validated. Batch-to-
batch segregation represents a
challenge to all therapeutic
manufacturing processes. For small
batch sizes, however, the impact of
conventional cleaning and sanitizing
strategies on operating costs and
facility uptimes can be significant.
The bioprocess industry has
recently seen an emergence of
presterilized, closed-consumable kits
and associated equipment. These have
been used for achieving product
isolation in conventional cell culturing
processes. As closed-consumable
technology has matured and gained
acceptance, the same philosophy has
also been applied to several upstream
and downstream processes such as
closed, continuous centrifugation.
KBI Biopharma Inc.s (www.
kbibiopharma.com) kSep technology
is one recent example that illustrates
the potential of closed processing
and how scalability can be
addressed. The closed, continuous
centrifuge was developed to gently
concentrate, capture, and/or separate
cells by providing a very low-shear
and nourishing environment. The
kSep technology is being applied in
a single-use format to take
advantage of shorter cycle times,
fewer validation requirements, lower
processing costs, and minimal risk
from cross contamination.
Photo 4 shows two kSep variants
based on an identical core process
technology but capable of
significantly different throughput
rates. Using them, development
work can be performed at small
scale with confidence: If or when
larger-scale batches need to be
processed, a bioequivalent solution
will be available.
As mentioned, closed-consumable
kits enable us to address the risk of
contamination during manual
operations such as sterile connections.
Such kits are also effective for
ensuring batch-to-batch segregation.
For cell therapies, each process is
unique. They typically involve
collection and isolation, then culture,
expansion, and manipulation of cells
followed by harvesting, washing,
filtering, and concentrating them;
formulation and product filling,
storage, and transportation; and
finally administration to patients.
Assuming that a cell therapy process
can be designed so that it remains
closed throughout, implications on
cost and quality can be profound.
Because a closed consumable is sterile,
associated risk of contamination from
its environment can be considered
negligible. On that basis and
provided that a consumable remains
closed throughout the entire process
processing can be undertaken in
lower-grade cleanrooms. It also means
that different batches (provided that
they too are closed) can be
concurrently processed in the same
room. This could significantly reduce
the need to build and operate
expensive cleanrooms.
To realize the full potential for a
cell therapy using closed-
consumable processing therefore
requires a closed solution for every
manufacturing and QC step in the
process, and herein lies the
challenge. Although closed
processing solutions now exist for
some process steps, they are often
not ideally suited to the
manufacturing requirements of a
specific cell therapy and thus are
not readily integrated.
Reengineering a cell therapy process
to make it closed and satisfy all
requirements for a specific cell
therapy requires
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of the process and its critical
parameters
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capabilities of available technologies
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gaps and development of novel
solutions
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user-friendly, error-free process that
can be quickly and reproducibly
scaled to match demand (Photo 2)
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requirements for CGMP
manufacturing.
CLOSED AND AUTOMATED
CELL PROCESSING
How It Works: Cell therapy processes
including centrifugation, incubation,
media addition, cell selection, cell
washing, and final fill and finish can
be performed within closed
consumables (Figure 2). Machines
integrate and automate the processes
and replicate many qualitatively
controlled manual tasks. They also
provide consistent and operator-
independent quality. Because
processing is closed, automated
machines producing therapies for
different patients can be placed side-
by-side in a processing room, whereas
conventional open processing would
require a separate sterile environment
for each patients therapy.
Benefits delivered from an
automated, closed-cell processing
system would include significant
reduction in the cost of therapies
(typically 2590%) and the number of
operators required (typically >70%);
lowered dependence on skilled labor;
Photo 5: kSep closed continuous
centrifuge machines
Photo 4: Loading Argos Therapeuticss
automated RNA machine with a
closed consumable kit
MARCH 2011 BioProcess International 9 SUPPLEMENT
significant savings in capital
investment through better facility use
(typically 3050%); improved quality
and fewer quality events; and an
ability to more rapidly scale up and
scale out to match market demands.
An integrated cell therapy
manufacturing facility (Figure 3)
comprises a central processing room
containing several automated
machines that process multiple
patients and batches in parallel.
Around that are situated other
facilities and processes needed to
support those operations including,
supply, storage, and retrieval of input
and output materials including
reagents, disposable processing sets,
patient materials, finished products,
and waste; collection and processing
of QC samples for data capture and
process feedback; and a process
recovery suite. This allows use of
specialized facilities to be maximized
and enables rapid scale-up of
production to match demand.
SCALE UP OR SCALE OUT?
My company has been involved in
developing several automated systems
for both autologous and allogeneic
cell therapy processes. Despite
differences in the types of cells and
processes required to produce these
therapies, the scale-up challenges
they face are remarkably consistent.
Cost: Cell therapy batch sizes are
often small (especially for autologous
products), so applying conventional,
large-batch processing strategies is a
poor use of facilities. Large GTP/
GMP cleanroom facilities are
impressive and glamorous, but very
costly to build, and even more
expensive to operate (3). Dendreon
Corporation (www.dendreon.com),
which received US FDA approval for
its Provenge prostate cancer treatment
in 2010, specifically describes some
risks to scale-up:
The costs of expansion of our
facilities and investment in related
equipment has been and will
continue to be a very significant
expenditure for us during 2010
and 2011. In order to
commercialize Provenge, in the
event of licensure by the FDA, we
will need to hire and train a
significant number of employees
and comply with applicable
regulations for our facilities,
which are extensive. In addition
to the monetary costs of
expansion of our manufacturing
capabilities, the facilities
build-out requires significant time
and attention of our executive
management (6).
Automation: Most cell therapy
processes have been developed in a
laboratory. As a consequence, they are
manual and labor-intensive, and they
require a high degree of skill. This may
be logical and manageable during
research, but it would be considerably
more difficult for commercial-scale
production. Processing technologies
typically used during clinical trials
arent often optimized or easily
integrated for cell therapy production.
In a CGMP production environment
where the product is the process,
change is only possible under very strict
controls. Changing manual processes
into more efficient, automated
processes after regulatory approval is
often difficult, time-consuming, and
expensive. In fact, we have seen several
cases in which cell therapy companies
are committed to technology that
they know is suboptimal or, worse still,
no longer available.
A trained researchers mindset is
forever locked into the quest for
innovation and improvements as well
as the drive to act on new findings
in short to experiment. Minds
normally dedicated to investigating
unexplored elements at the forefront of
medicine would need to switch gears
to engage in operating a production
process that is reproducible, consistent,
and invariably repeatable. This
fundamental mismatch in mindset
would be a waste of talent and
training. Companies embarking on a
route to scale-up, unsurprisingly,
report worker dissatisfaction and
significant difficulties in recruitment
and retaining of staff.
Risk: The potential of cross
contamination and/or loss of patient
identity (chain of custody) for each
batch must be maintained at a zero
risk level as scale-out occurs (1).
Apart from the inherently high cost
of manual cell therapy processes, it is
therefore extremely difficult to
imagine how you could efficiently
scale up or scale out a manual process
without significant deviations. Up to
50% of batch failures are attributable
to operator error, and it is intuitive
that the increasing complexity of an
operation raises the likelihood of
errors occurring (1).
The primary strategy that
Invetech has applied to small-batch
processing is to create a new type of
factory in which manufacturing
processes are closed and the most
complex and vulnerable process
steps automated. With careful
design and a real-time quality
control system that detects
erroneous states before process
Figure 4: Typical development stages for cell therapies with closed, automated processing
IND
10 patients
Phase 1
10 patients
Phase 2
20 patients
Phase 3
50 patients
Commercialization
100 patients 1,000 patients 10,000 patients
Plan Develop, Prove
Roll Out
10 BioProcess International MARCH 2011 SUPPLEMENT
deviations are created, a
manufacturing process can thus be
created that achieves a commercially
viable cost of goods and eliminates
critical failure modes.
The system developed by Invetech
for Argos Therapeutics represents a
previously manual process that was
successfully redesigned based on the
guiding philosophy of closed-
consumable processing on an integrated
and automated production machine.
We developed an automated cellular
and RNA processing system to
manufacture autologous
immunotherapies for treating advanced
kidney cancer (metastatic renal cell
carcinoma), B-cell chronic lymphocytic
leukemia, and HIV infection. At the
heart of the automated process are
three standalone units: an RNA
processing subsystem (Photo 4) that
processes tumor homogenate into
amplified tumor RNA, with all steps
performed on a single patient sample
and taking place in a closed disposable
container; and two cellular units
designed to perform a series of cellular
and plasma processing steps to generate
a final product in functionally closed
disposable vessels.
The system removes complex and
vulnerable manual operations and
replaces them with automated
processing steps that can be
consistently reproduced. Each unit can
be validated and will perform precisely
the same function no matter where in
the world it is operating. This
substantially eliminates the most
significant contributor to variation:
operators. We now have a process that
can be scaled out to match demand by
installing additional, automated
machines in either the original facility
or in others that are located closer to
potential markets. By locking down
the process with automation and
closed consumables, we have also
significantly reduced the companys
reliance on hiring and training skilled
operators as well as the cost and time
required to establish dedicated
cleanrooms.
ROADMAP TO COMMERCIAL-
SCALE PRODUCTION
Cell therapy companies need to invest
in alternative technologies to ensure
that they can have commercially viable
products. However, it is also
important to recognize that funds are
typically precious during early stage
clinical trials. Until a therapy is
proven to be safe and efficacious, and
a reasonably mature process exists,
investment in highly customized
systems should be kept to a minimum.
But after phase 3 clinical trials using a
specific process are complete, changes
to that process can be extremely
difficult to make and will require
proof of bioequivalence or even
additional clinical trials.
Developing a customized CGMP
system will typically take two to four
years, depending on the complexity of
the process and the time it takes to
progress through development, clinical
trials, and capital raising. Invetechs
recommendation is to invest a
relatively small amount of effort and
money at phase 1 to identify the most
appropriate scale-up system and plan a
development strategy that includes
cost and timing (Figure 4). Another
primary objective of the feasibility and
planning stage is to identify core
technologies that can be used for
initial trials and subsequently scaled
up and integrated into a CGMP
commercial-scale production system.
Ideally, development of a
commercial production system should
commence during phase 1 or 2,
depending on the development time
scale and clinical trial schedule. At a
minimum, the key objective should
be to have a commercial-scale system
available during phase 3. This
ensures that those pivotal trials are
conducted using a process that is
equivalent to the commercial
production process, which prevents a
need to make expensive and time-
consuming postapproval changes.
Photo 6 shows one of the outputs
of such a feasibility study that my
company undertook for MolMed SpA
before commencement of phase 3
clinical trials for its TK therapy.
The product is an ex vivo cell therapy
to enable safe haematopoietic stem cell
transplants (HSCTs) from partially
compatible bone-marrow donors to
treat hematological malignancies,
particularly high-risk acute leukemia.
Debate will continue as to the
relative merits and likelihood of success
for allogeneic and autologous cell
therapies. Ultimately, the overall
commercial success of any such product
will depend on it being safe,
efficacious, cost-effective, and scalable.
It also needs to be affordable and
accessible to a substantial patient
population. For some companies,
allogeneic cell-based therapies are
attractive because they reflect the
current large-batch pharmaceutical
investment model and have been
supported on this basis. However, not
all allogeneic therapies would be
considered large-batch products. Many
autologous therapies could provide
significant therapeutic advantage.
At Invetech, we believe that for
the cell therapy industry to reach its
full potential, it must have
commercially viable small-batch
processing and ideally the ability
to deal with a batch size of one
(patient). Some cell therapy
Photo 6: A closed and automated system
from a feasibility study undertaken for the Italian
company MolMed SpA (www.molmed.com)
C
For the cell therapy
industry to reach its
full potential, it must
have commercially
viable small-batch
processing and
IDEALLY
the
ability to deal with a
batch size of one.
companies have already invested in
developing innovative closed,
automated systems for small-batch
cell therapy manufacturing. Those
systems have delivered significant
cost, quality, and scale-up benefits
over existing manual, cleanroom-
based processing strategies. They
have also provided valuable insight
into the industrys needs and
demonstrated what is possible.
Cell therapy no doubt is an
extremely exciting and rapidly
advancing field of medicine that
could have a significant impact on
how we treat a vast array of diseases.
To reach its full potential, however,
the technology will require more
than old-fashioned factories. Closed
and automated cell-processing
systems provide an opportunity to
change the paradigm and deliver
these therapies at a cost that was not
previously thought possible.
REFERENCES
1 Mason C, Dunhill P. Assessing the
Value of Autologous and Allogeneic Cells for
Regenerative Medicine. Regen. Med. 4(6)
2009: 835853; www.slv.vic.gov.au.
2 How Do Drugs and Biologics Differ?
Biotechnology Industry Organization:
Washington, DC, 2010; www.bio.org/
healthcare/followonbkg/DrugsVBiologics.asp.
3 Burger SR. Design, Operation and
Management of GTP/GMP Cell
Engineering Facilities. BFDA 2007
International Symposium on Regulation of
Human Cell- and Tissue-Based Products.
Taipei, Taiwan, 3 October 2007; www.ac-gt.
com/talks/Dec 2010.
4 Shaw R. Industrializing Stem Cell
Production. BioProcess Int. 8(9) 2010: 1015;
www.bioprocessintl.com/journal /2010/
October/Industrializing-Stem-Cell-
Production-303899.
5 Hampson B, Rowley J, Venturi N.
Manufacturing Patient-Specif ic Cell
Therapy Products. BioProcess Int. 6(8) 2008:
6072; www.bioprocessintl.com/
journal /2008/September/Manufacturing-
Patient-Specif ic-Cell-Therapy-
Products-183198.
6 Form 10-K Annual Report Pursuant to
Section 13 or 15(d)of the Securities Exchange
Act of 1934. Dendreon Corporation: Seattle,
WA, 31 December 2009; http://f iles.
shareholder.com/downloads/
DNDN/1125257529x0x420918/E230B7F2-
3912-4F73-AD30-9208DA7DBA53/
ar_2009.pdf.
7 CDER/CVM/ORA. PAT Guidance for
Industry: A Framework for Innovative
Pharmaceutical Development, Manufacturing,
and Quality Assurance. US Department
of Health and Human Services, Food and
Drug Administration: Rockville, MD,
September 2004.
FURTHER READING
Mason C, Hoare M. Regenerative
Medicine Bioprocessing: Building a
Conceptual Framework Based on
Early Studies. Tiss. Eng. 13(2) 2007;
www.slv.vic.gov.au Nov 2010.
C
David James is director of life science
and pharmaceutical manufacturing
innovation for Invetech Pty. Ltd., 495
Blackburn Road, Mount Waverley,
Victoria 3149, Australia; 61-3-9211-7742;
david.james@invetech.com.au,
www.invetech.com.au.
To order reprints of this article,
contact Carmelita Garland (carmelitag@
fosterprinting.com) at 1-800-382-0808, ext.
154. Download a low-resolution PDF online
at www.bioprocessintl.com.
n
e
t
h
e
p
o
GE Healthcare
Life Sciences
GE, imagination at work and GE monogram are trademarks of General Electric Company.
WAVE Bioreactor and Ficoll-Paque are trademarks of GE Healthcare companies.
All third party trademarks are the property of their respective owners.
2011 General Electric Company All rights reserved. First published Feb 2011
GE Healthcare Bio-Sciences AB, Bjrkgatan 30, 751 84 Uppsala, Sweden
GE Healthcare at the ISCT Meeting in Rotterdam
May 23 25, 2011
Visit our booth for a demonstration of automated expansion of cells in suspension using
the WAVE Bioreactor
System 2/10
Also featuring
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38 BioProcess International MARCH 2011 SUPPLEMENT
CELL THERAPI ES TECHNOLOGY
Meeting the Challenges in
Manufacturing Autologous
Cellular Therapies
by Tamara T. Monesmith
P
ersonalized medicine is a
promising new approach to
disease treatment. The
ultimate in personalized
medicine is a cellular therapy
manufactured specifically for an
individual patient using his or her own
cells. But this autologous approach to
generating immunotherapies has
unique manufacturing challenges.
Each patient receives an individual
product batch, which needs to be
manufactured, tested, and released. So
thousands to tens of thousands of
batches could be made for each
indication every year. Given the
personalized nature of these therapies,
the production scale remains the same
for each batch. Thus, scale-up is not
required; scale-out is key for meeting
the demands of autologous cell-
therapy manufacturing.
MANUFACTURING METHODS
Generating autologous cellular
therapies can take several approaches.
In general, all involve obtaining
autologous cells from a patient by
procedures performed at a hospital or
blood center. Depending on the
therapy, those cells are either
transferred to a local laboratory for
further processing or shipped to a
central processing facility. This
starting material has limited stability,
requiring immediate processing upon
receipt. For proliferating cells, the
desired cell type for a given therapy is
typically expanded in cell culture. If
the cells are terminally differentiated
(nonproliferating), then cells for the
therapy are typically generated by
isolating precursor cells for the desired
type and culturing them with
appropriate cytokines for
differentiation into the desired cell
type.
During cellular processing, an
antigen may be added to elicit a
desired immune response. This may
be a defined antigen (the same for
every patient and process) that can be
produced using traditional large-scale
methods. For example, Dendreon
Corporation uses a defined-antigen
approach in making its Provenge
(sipuleucel-T) prostate cancer
treatment, which received the first and
only FDA approval to date for an
autologous cellular therapy in April
2010. The antigen for Provenge is a
recombinant fusion protein comprising
granulocyte macrophagecolony
stimulating factor (GM-CSF)
crosslinked to the prostate antigen
prostatic acid phosphotase (PAP).
This recombinant fusion protein
(PAP-GM-CSF) is manufactured at
large scale to provide material for
multiple batches of product.
Other manufacturing approaches
use an autologous antigen from each
patients tumor for oncology
treatment or from a given virus for
treating infectious diseases. They
generate completely autologous
cellular therapies tailored to each
individuals disease. This enables
adaptation of methods from one
indication to another without the
need to identify defined antigens for
each indication. However, it also
increases the complexity of the
manufacturing process because an
antigen needs to be processed and
tested for each patient and combined
with that patients own cells.
There are different methods for
processing autologous antigens. The
simplest method is to use tumor lysate,
which requires relatively minimal
processing, essentially a
homogenization step. That provides a
Photo 1: Argoss prototype device for amplified
RNA processing was developed with the
Australian company Invetech Pty. Ltd.
patients cells directly with the proteins expressed by his or
her tumor as antigens during cellular processing to generate
the immunotherapy. Alternatively, tumor lysate can be
processed to isolate total RNA, enabling cells of interest to
express and process it for the immunotherapy. This can be
taken a step further by amplifying the mRNA in the
isolated total RNA. Although that requires the most effort
in terms of antigen processing, a relatively small amount of
tumor is required to generate sufficient mRNA for multiple
doses (or even multiple batches) of the cellular therapy.
Following cellular processing, the resulting cellular
therapy may be either directly infused (so one
manufacturing run generates one dose for the patient) or
cryogenically preserved (in multiple doses). Once
cryogenically preserved, each individual dose for a patient
is then delivered using liquid nitrogen dry shippers before
administration.
In early phase clinical trials, the manufacturing of
cellular therapies generally uses manual processing methods
transferred from research laboratories. Semiautomated
commercial equipment options are available for the
isolating precursor cells. Some instruments use antibody-
coated beadbased methods (e.g., the CliniMACS cell
separation system from Miltenyi Biotec, www.
miltenyibiotec.com) for isolating precursor cells. Another is
based on elutriation, also known as counterflow
centrifugation (the Elutra cell separation system from
CaridianBCT Inc., www.caridianbct.com). Some
companies have developed their own proprietary methods
for isolating precursor cells. Examples include a tangential-
flow filtration method developed by Northwest
Biotherapeutics, Inc. (www.nwbio.com) and a cell
separation device used by Dendreon.
Options abound for the initial isolation of precursor
cells. But little automated commercial equipment is
available to address either the remaining cellular processing
steps or the more complex antigen processing steps. Some
companies are working to develop cellular therapy enabling
devices, such as Miltenyis CliniMACS Prodigy system.
But a need remains for automated equipment that can
handle the complexity of autologous therapy processing and
enable scale-out for larger clinical trials and
commercialization. Therefore, Argos Therapeutics initiated
development of its own automated system to address these
manufacturing challenges.
The key to scale-out for autologous cellular therapies is
using functionally closed, single-use technologies and
automation for processing. Such disposables minimize
cleaning requirements between batches which is critical
for turnover of equipment and eliminate concerns
regarding cross contamination. Automation provides
consistency and efficiency in processing using disposables.
A CUSTOM DESIGN
Argoss Arcelis platform generates monocyte-derived
dendritic-cell products in clinical development for
oncology and infectious disease. The technology uses
amplified RNA from a patients tumor sample as an
Ccscd SysLcns rcn Ccrnng
ln Ihe race Io bring new drugs, Iherapies and vaccines Io markeI, researchers
rely on innovaIive Iools IhaI compress cosIs and Iimelines. Corning's leading-
edge qualiIy bioprocess producIs also oIIer Ihe opIion oI closed sysIems
which improve producIiviIy, enabling your breakIhrough biopharmaceuIical
producIs Io go speedily Irom k&D Io markeI.
WaIch our RPLk video aI www. corning.com/Iifesciences/whatsnew.
www.corning.com/Iifesciences
40 BioProcess International MARCH 2011 SUPPLEMENT
products in clinical development for
oncology and infectious disease. The
technology uses amplified RNA from
a patients tumor sample as an
antigen (Figure 1). In collaboration
with Invetech Pty. Ltd. (www.
invetech.com.au), Argos is developing
automated equipment that uses
functionally closed disposables to
enable commercial manufacturing.
White blood cells obtained from
patients through leukapheresis are
shipped to the Argos central
manufacturing facility for processing.
Isolation of monocytes (the precursor
cells used to generate dendritic cells)
is performed using a functionally
closed disposable and the existing
semiautomated Elutra cell separation
system. Two additional devices are in
development to perform the
remaining cellular processing steps:
media-exchange steps required to
prepare the cells for culture,
electroporation (the process used to
introduce antigen RNA into cells),
and formulation and fill of the drug
product. Because the drug product
formulation uses each patients own
plasma as an excipient, one device
processes the plasma, which is
collected during leukapheresis, and
prepares it for use in the final
formulation of the cellular therapy.
Patient material never comes into
contact with the equipment, only the
disposable, which prevents cross
contamination and minimizes
cleaning between processes. The use
of tube sealers and tube welders
enables movement (removal or
addition) of bags or reagents as
required. For example, when culture
bags are filled with the appropriate
cells, media, and cytokines, they are
removed using a tube sealer to be
incubated for the required five days.
When cells need harvesting, those
culture bags are attached to the
appropriate disposable using a tube
welder. Methods are in place to verify
that the appropriate patients cells
will be welded onto the disposable.
Similar methods verify that the
correct tubes are being welded before
welding is enabled for all reagents
and in-processing materials.
Argos and Invetech designed a
third prototype device (Photo 1) to
process tumor homogenates to isolate
and amplify RNA inside a
functionally closed disposable
container. This single-use technology
has been uniquely designed to ensure
that equipment does not come into
contact with patient material (Figure
2). Again, this eliminates concerns
for cross contamination between
processes. It also eliminates the need
for cleaning validation, which would
be extremely challenging because the
product generated during this process
is amplified nucleic acid.
The disposable containment
consists of a rigid tray incorporating
a PCR plate for nucleic acid
amplification. It also contains
washing and elution stations for
isolation and purification steps. A
disposable pump head is
incorporated to generate a closed-
loop vacuum for isolation and
purification using silica columns. An
incorporated reagent rack can be
cooled to ensure reagent stability.
All necessary reagents are foil-sealed
until required for processing. There
are tip racks to hold new and used
pipette tips for all liquid transfers. A
f lexible barrier top with an
incorporated pipette head is sealed
onto the rigid tray. The barrier
allows a pipette head to access all
areas inside the tray: tip racks,
reagent racks, PCR plate, and
washing/elution stations.
The device contains analytical
equipment needed to determine
concentration of the nucleic acid,
with spectrophotometric cuvettes
incorporated in the rigid tray for
such testing. Volumetric cuvettes are
designed to enable determination of
the volume of resulting nucleic acid
solutions generated to calculate
yields and volumes required for
concentration normalization. The
thermal cycler lid moves through the
side of the disposable through a
rolling boot, enabling it to move in
and out to cover the PCR plate
during incubations and allowing
pipette access for transfers to and
from the plate. Overall, this
equipment and disposable design
provides a fully automated method
for RNA isolation and amplification
from tumor homogenate in a single-
Figure 2: Argoss single-use technology for functionally closed processing of amplified RNA
Flexible barrier to allow
robot arm movement
Disposable vacuum
pump head
Pipette head attached
to robot arm
Thermal cycler lid inside
thermal cycler boot
Sealed tubing for aliquots
Figure 1: Argoss Arcelis platform for generating monocyte-derived dendritic cell therapies
Clinical Site Argos Therapeutics
Small amount of
tumor/pathogen
Leukapheresis
Intradermal injection
Cryoshipper
Total RNA
Amplifed RNA
Monocytes
Mature Dendritic Cell
Vial and Freeze
Mature, Electroporated
Dendritic Cell
use format. This is critical for
meeting the demands of commercial
manufacturing for Argos
Therapeutics.
MORE CHALLENGES
In addition to the need for automated
processing equipment, other critical
considerations come
up when processing autologous cellular
therapies on a commercial scale. The
need to process the patients cellular
samples immediately upon receipt of
them makes knowing when a therapy
is prescribed and scheduling of their
collection critical. If an antigen is
autologous as well, then companies
must ensure that material is received,
processed, and ready to be
incorporated into cellular processing.
Given the unique traceability
requirements for autologous cellular
therapies, mechanisms to ensure that
multiple patient product streams are
combined correctly and that the
correct product is delivered to each
patient are essential. The
extraordinarily high number of
batches being tested and released for
commercialization needs to be
addressed. The complexity of process
scheduling, traceability, and release
issues for autologous cellular therapies
requires a well-designed
manufacturing execution system to
manage those activities and enable
electronic batch records with release
by exception. For the promise of
personalized cellular therapies to be
fully realized, effective methods need
to be developed that will address all
these challenges and complexities of
their manufacture.
FOR FURTHER READING
Goldman B, DeFrancesco L. The Cancer
Vaccine Roller Coaster. Nat. Biotechnol. 27(2)
2009.
Nicolette C, et al. Dendritic Cells for
Active Immunotherapy: Optimizing Design
and Manufacture in Order to Develop
Commercially and Clinically Viable Products.
Vaccine 25, 2007: B48B60.
Whiteside TL. Evaluation of Dendritic
Cell Products Generated for Human Therapy
and Post-Treatment Immune Monitoring.
BioPharm Int. 21(3) 2008: 4267.
Miesowicz f. Dendritic-Cell Based
Therapies. Genetic Eng. News 27(20) 2007:
4446.
Finke LH, et al. Lessons from
Randomized Phase III Studies with Active
Cancer Immunotherapies: Outcomes from the
2006 Meeting of the Cancer Vaccine
Consortium (CVC). Vaccine 25S (2007): B97
B109
Osada T, et al. Dendritic Cell-Based
Immunotherapy. Int. Rev. Immunol. 25, 2006:
377413.
Hoos A, et al. A Clinical Development
Paradigm for Cancer Vaccines and Related
Biologics. J. Immunother. 30(1) 2007: 115.
Kantoff PW, et al. Sipuleucel-T
Immunotherapy for Castration-Resistant
Prostate Cancer. N. Engl. J. Med. 363, 2010:
411422.
Fitzpatrick I. Cellular Therapy Success
Through Integrated Automation. BioProcess Int.
6(9) 2008: S32S37.
Rios M. Flexible Manufacturing. BioProcess
Int. 8(5) 2010: 3446.
C
Tamara T. Monesmith is director of
manufacturing and process development at
Argos Therapeutics, 4233 Technology Drive,
Durham, NC 27704; 1-919-287-6321, fax 1-919-
287-6301; tmonesmith@argostherapeutics.
com, www.argostherapeutics.com.
To order reprints of this article, contact
Carmelita Garland (carmelitag@
fosterprinting.com) at 1-800-382-0808, ext.
154. Download a low-resolution PDF online
at www.bioprocessintl.com.
42 BioProcess International MARCH 2011 SUPPLEMENT
CELL THERAPI ES COMMERCIALIZATION
Industry Roundtable
Viewpoints on Processing, Quality, and Regulations
by Maribel Rios
W
ith one eye on
commercialization and the
other on monitoring every-
day challenges, cell therapy
manufacturers are asking critical
questions about process efficiency,
ensuring quality, and satisfying
regulatory demands. In this virtual
roundtable discussion (participants
were asked questions separately), cell
therapy industry representatives
answer key questions in hopes of
broadening understanding about this
new class of biopharmaceuticals.
Participants in this roundtable are
Timothy Fong, PhD (director cell
therapy, Becton Dickinson
Biosciences), Annemarie Moseley,
PhD, MD (CEO, Repair
Technologies), Firman Ghouze
(director of cell therapy, GE
Healthcare), Aby Mathew, PhD
(senior vice president and chief
technology officer, BioLife Solutions),
and Robert Deans (vice president of
regenerative medicine at Athersys and
ISCT committee chairman).
PROCESSING
BPI: How does the processing of
therapeutic cells differ from that for
traditional biologics?
Fong: Cells are rather fragile, so
techniques used for purifying small
molecules or biologics are too harsh,
because they are mostly physical
methods of isolation with potentially
extreme non-physiological conditions.
Many cell processing protocols use a
combination of traditional cell culture
methods and techniques used in
peripheral or cord blood banking such
as elutriation. When I think about cell
therapy processing, I think more about
antibody-based separation methods.
There are two main types: magnetic-
beadcoupled antibody reagents and
flow-activated cell sorting (FACS).
However, as of today no FACS systems
are what I would consider to be
completely compliant with current
good manufacturing practice (CGMP).
So cell processing companies need to
jump through some additional hoops in
terms of process development and
validation to comply with regulatory
aspects of cell CGMP processing.
Moseley: If the cells are frozen, they
need special handling, and you need to
pay attention to the timing of
therapies. Under biotechnology
conditions cells cannot survive or they
are modified. Cell therapy processing
reactors are different, and there are no
ultracentrifuges. Currently there is not
even a good way to reduce volume from
a large cell collection. So there is a lot
of room for manufacturing
improvements all likely to come
after the first product is approved.
Ghouze: Some manufacturers use
tangential flow filtration (TFF), in
which the cells arent rammed against a
filter membrane. Many facilities that
develop cells either have discrete closed
systems to process these cells, or they
have a large CGMP-type laboratory
that works with cells.
Fong: One of the current major
needs in processing and isolating cells
in general is the development of better
closed systems in which we can isolate,
grow, activate, and expand them. The
technology is emerging, but we dont
have a lot of choices in hand to
facilitate that.
Moseley: Manufacturing is still very
people-dependent. It requires a lot of
oversight and very intense
manipulation by personnel, as opposed
to robotics and automation in
biopharmaceutical processing. Scale-up
and logistics are still in their infancy.
Its much like the antibody industry, in
which initially no one could do it, so
companies had to hire contractors.
Now everyone is making antibodies
in-house. I think that is eventually
where we are going to see this field
moving.
BPI: What equipment is used to
manufacture cell therapies?
Moseley: Most companies have
customized processes, so there are no
standardized bioreactors, for example.
Cells used in antibody production have
been manipulated to the point where
they can be in suspension, but most
Human mesenchymal stem cells
preserved for three days with
HypoThermosol FRS media from BioLife
Solutions WWW.BIOLIFESOLUTIONS.COM
MARCH 2011 BioProcess International 43 SUPPLEMENT
adult stem cells like being attached to a
surface. Designs using many tubes or
beads have been tried, but right now
there are only a few containers that
have a lot of surface area. The primary
of those are Cell Factories (Nunc,
ThermoScientific, www.nuncbrand.
com). They resemble a lot of tissue
flasks glued together. Those designs
have not addressed scale-up. They are
simple because so far there hasnt been
much demand for them. As demand
increases, the industry will need a cell
reactor. But right now, there arent
enough companies with products in the
intermediate phases, so they are having
to adapt their collection systems and
centrifugation steps.
BPI: How will scale-up become an issue
as products progress to commercialization?
Ghouze: Getting cells of the
required quality in a scalable manner
will be a challenge. Companies can
successfully process them at a scale for
small clinical studies (trials with 1020
patients). But if some of those cell
therapies progress in the way industry
hopes, and if clinical trials progress
well, then companies will need scalable
solutions from a CGMP facility to
enable really widespread distribution.
Moseley: Each Cell Factory system
has a few liters of volume media in it
that must be centrifuged to collect the
cells, which is a problem at scale-up.
Currently there are no large-scale
systems for handling that much media.
So researchers have been trying to
figure out different ways to speed up
the process of centrifugation before
cells are diluted in freezing media. All
the centrifuge systems used to collect
the antibodies off of the reactors are
way too harsh for the cells because
antibodies are so much smaller. The
process of getting massive volumes of
cells and combining them is not
automated, and companies have been
trying to figure out ways to do that
efficiently. There are some real
opportunities for process development,
automation, and new devices. Its just a
question of what is going to trigger that
work. A lot will depend on market
need and whether many companies will
be doing that part of the process
themselves or whether it will be done
through a contract services provider.
BPI: Cryopreservation is an important
processing step for some cell therapies.
What techniques are currently used?
Mathew: Techniques are usually
separated between the standard
cryopreservation methods (used by
most) of traditional ice management
home-brew freeze cocktails or the
newer class of intracellular like complex
formulations; and then to a lesser
degree vitrification. Certainly,
controlled-rate freezers have allowed
programming and greater manipulation
of freezing rates throughout a
cryopreservation protocol, but even
isopropanol freezing containers have
made it easier for folks to have more
consistency in their cryopreservation
protocol.
BPI: What tests are used to ensure cells
are viable after cryopreservation?
Mathew: All biologics are normally
maintained under normothermic
conditions either within the body or as
part of cell culture conditions.
However, once cells are removed from
normothermic conditions, changes
occur in their metabolism and cellular
integrity. If these changes are
reversible, then cells can recover
appropriately upon return to normal
conditions. If the stresses of these
changes build up too much, the cell is
negatively influenced to undergo active
pathways of breakdown (such as
apoptosis) or passive pathways of
breakdown (such as necrosis).
Tests used to ensure viability
(and/or functionality) are varied and
debatable regarding appropriateness.
For cell therapy, viability is often
assessed immediately postthaw by
simple live/dead assays that may not
indicate the true, long-term viability of
BPI: What is your understanding of FDA
requirements for assays and testing?
Moseley: The FDA requires the same
global release criteria in terms of sterility,
potency identity, and so forth as for
antibodies. Assays to date have been
based on the identity of the cells as stem
cells, and potency assays have varied
based on the indication and whether a
company chooses to focus on indication
potency or potency as to stem cellness.
The same types of toxicity testing apply,
although biodistribution is different and
duration of the studies has been longer
in some cases, making preclinical studies
more costly.
BPI: What is your understanding about
guidelines for operator safety?
Fong: Peripheral and cord blood banks
primarily use closed cell-processing
systems. A Class 10,000 room is perfectly
acceptable for those types of systems.
Other processes such as FACS cell
isolation are not closed systems. My
current understanding is that a Class
10,000 room or equivalent is probably
required in addition to some sort of
biosafety cabinet for flow instruments.
Obviously, a major concern is operator
safety because flow sorters are not
closed systems, and most of them have
some level of aerosolization. As far as
handling of large numbers of cells
before, during, and after cell
manipulation, a lot of the systems that
cell therapy developers are using today
have been co-opted from the blood
banking industry. So suppliers need to
think how they can develop more
specific closed systems for all the
different types of cells stem cells,
adherent cells, suspension cells that
companies will be wanting to grow for
future cell therapies.
A lot of developers have become very
familiar with the guidance for cord blood
banking, tissue processing guidelines,
and biologics manufacturing guidelines,
particularly when it concerns
manufacturing reagents. Take antibody-
based separation methods, for example.
If you look through the FDA guidelines
for cell therapy, it refers back to
manufacturing guidelines for therapeutic
monoclonal antibodies. So in a sense,
even though your antibody reagent is
really part of your process for isolating
your cells, manufacturing to guidelines is
essentially equivalent to a therapeutical
monoclonal antibody. Having said that, I
believe that most regulatory agencies are
flexible, and theyre looking at each IND
application and cell manufacturing
process case by case to determine
whether the level of CGMP compliance is
acceptable for that particular application.
As far as I can tell, in general, FDA and
EMA are trying to follow similar
guidelines. One issue unique to Europe is
that in addition to EMA regulations, you
have national and regional regulations
that can add another layer of complexity.
REGULATIONS AND SAFETY
44 BioProcess International MARCH 2011 SUPPLEMENT
the cells due to the phenomenon of
preservation induced, delayed onset
cell death.
During cell therapy product
characterization and manufacture, it
would behoove a manufacturer to
validate the efficacy of its preservation
methods with multiple viability,
potency/functionality, and stability
assays. For broad application to many
cell types, you can use tests such as
live/dead assays that can be assessed
with a fluorescent plate reader or flow
cytometer. Metabolic assays and
membrane integrity tests are another
method of assessing cell health.
Furthermore, analysis of the -omics
(e.g., genomics, proteomics,
metabolomics) allows for in-depth
characterization of a cell product. And
these assessments should be performed
on the cell product at multiple stages in
the stability intervals to better
understand the limitations of the
overall system of manufacturing,
banking, and/or transport, and clinical
delivery application. An optimized
biopreservation system would allow
clinical and commercial development of
cell and tissue products with the
greatest clinical efficacy, lowest
manufacturing costs, and greatest
stability to enable worldwide
distribution.
The biggest disparity is that there is
the academic/basic science
perspective to cryopreservation and a
process development view.
Academically, each cell/tissue type has
its own characteristics of water content,
membrane permeability, size, sensitivity
to stress, and so forth. However, from a
process perspective you do not want to
have to use a custom process and
custom solution for each cell type. A
cell therapy company that invests a lot
into its specific cells (such as custom
growth media) may be okay with a
custom freeze process if all it is
working on is that cell type. But if you
are a cell supplier, or a pharmaceutical
toxicity group that might be
concurrently working on CHO cells,
hepatocytes, or iPS cardiomyocytes,
then no one will want to have to use a
separate cryopreservation process for
each of these cell models. Cell therapy
groups do not want to invest more time
and resources than needed into
optimizing freeze media or cooling
rates, so cryopreservation methods that
support a streamlined, consistent, and
effective process will enable them to
achieve their goals sooner rather than
later.
BPI: Are there concerns about safety and
quality?
Moseley: Preclinical and safety
issues mostly refer to embryonic stem
cells. What you need to do to
demonstrate safety for the adult stem
cells has become almost routine now
with the FDA because probably up to
10 companies or more and many
protocols (over 50 already) have gone
through the FDA on this area. So that
is not so much an issue, but every
embryonic program will have its own
issues and programs.
Fong: One major misconception is
that cell therapy is a brand new field. If
you define cell therapy as using cells to
try to treat a medical condition, the
actual history goes back over 100 years
with blood transfusions and almost 60
years with bone marrow transplant.
However, what is new is the type of
cells and the sources of cells that we are
using. New developments are our
greater understanding of stem cells and
mature cells such as T cells and other
lymphocyte populations. We have a
better understanding of their
phenotypes, a better methodology of
isolating them to greater homogeneity,
and a greater understanding of how
cells act in normal physiology as well as
in disease. So now we can begin to
manipulate them and use them in our
favor to treat disease. That goes hand
in hand with improvements in cell
characterization and isolation.
Ghouze: Because bone marrow
transplantation has been around for a
long time, cell therapies based on bone
marrow cells have quite a robust quality
control (QC) process. There are various
techniques people can use to define
whether the cells used with
engraftment have a high likelihood of
success, for example. There is a drive
toward standardization, for QC
technologies to define what makes a
cell effective and safe. The industry is
currently moving in that direction, but
there is still some ways to go in that
area. The mechanism is poorly
understood in some instances. Having
said that, people will use existing
technologies such as flow cytometry,
but it must be scalable.
Robert Deans: There will always be
different perspectives on safety. With
the development of new and more
sensitive safety testing, tools, and
procedures, there is always going to be
a need to further educate and
reevaluate. But I dont see the adult cell
therapeutic space as being considered
highly risky at this point. Regulatory
agencies are encouraging development
and supporting trials in this area; and
there has been a very good safety track
record, especially in the case of
mesenchymal stem cells (MSCs). I
think there are now more than 5,000
patients who have been treated with
MSCs. So I dont see it as a safety
issue; I see it as a matter of education.
Potency assays are a very hot topic.
This relates to how cells are
characterized either phenotypically or
functionally, which in turn relates to
their biological performance. Part of
the issue also involves the intellectual
property that protects many cell
therapeutics. Investment and
development in this space is also going
to be driven around intellectual
property boundaries. We have an
insufficient hold on quantitatively
measuring potency and then comparing
potency among alternative but related
cell types. That is a very big area for
scientific and technology development,
and its one of the topics that is very
important for open debate so that
potential financial investors have an
opportunity to hear scientists make an
opinion on assays that they trust and
information they would require before
making a decision.
C
Maribel Rios is managing editor of
BioProcess International, 1-646-957-8884,
mrios@bioprocessintl.com.
To order reprints of this article,
contact Carmelita Garland (carmelitag@
fosterprinting.com) at 1-800-382-0808,
ext. 154. Download a low-resolution PDF
at www.bioprocessintl.com.
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CELL THERAPI ES BUSINESS
Addressing Business Models,
Reimbursement, and Cost of Goods
by Dawn Driscoll
T
he early ISCT organization
provided a powerful forum for
sharing solutions, developing
standards, and moving the
emerging concepts in cell therapy
forward as the field grew up and out of
academia. Currently, the ISCT
organization is uniquely positioned to
facilitate sharing of best practices,
standards, and strategies across the for-
profit cell therapy industry through its
Commercialization committee.
The Business Models,
Reimbursement and CoGS (cost of
goods sold) subcommittee of the ISCT
Commercialization committee was
formed to address several key business
topics with direct impact on the
industrys ability to develop, register,
and ultimately market cell therapies
successfully. The subcommittee aims
to define the issues as well as examine
successful approaches and solutions
companies have used, with a goal of
sharing best practices and strategies.
Activities in the United States, the
European Union, Japan, and China
will be evaluated, as well as in other
Asia-Pacific countries such as
Singapore and Australia. Although the
latter are smaller markets, they have
very well developed and advanced cell
therapy infrastructures and regulatory
environments.
CELL THERAPY BUSINESS MODELS
The business aspects of developing
and successfully marketing cell
therapies are genuinely complicated
but becoming better defined as the
field progresses.
An ideal cell therapy product
business model fits well with a
companys overall business plan and
integrates all relevant aspects of the
product throughout its lifecycle. Many
companies are currently wrestling with
overall go/no-go investment decisions
for cell therapy programs. Taking into
account all of a technologys
components can help determine the
appropriate manufacturing approach
and help determine whether that
technology can be a good fit within a
companys portfolio, and therefore a
solid investment.
As conventional wisdom has it, if a
technology is autologous, then its
primary emphasis will be on a service
model, whereas an allogeneic products
model is akin to a pharmaceutical,
long-termstorage model. However,
only taking allo- or auto- into
account is not sufficient for building
an appropriate business model for a
cell therapy.
All cell therapy business models are
dictated primarily by the technology:
e.g., the specific cell type, source
tissue and manufacturing process,
route of administration, and the
medical condition for which the
product will be used. Beyond that,
however, is the need to consider a
products market, including all current
and forthcoming competition; the
practice of medicine for that
indication; and flow of product,
patients, and money through the
overall system.
After those factors have been
considered, a company can build a
fully integrated forecast of production
(for example, the number of products
per month) for the entire lifecycle
(phase 13, launch, growth, and
maturity). This forecast will quantify
the needs for GMP manufacturing,
show when those needs occur, and
therefore help make the difficult rent
or buy (deciding between contract
manufacturing and internal
production) decision. Here is a
suggested pathway for mapping out an
overall model.
First, consider your product
technology, which encompasses the
following questions.
Source Tissue/Cells: Common or
rare? Hardy or delicate? Immunogenic
or not?
Production Process: Simple or
complex? Public or proprietary? Cheap
or expensive? Bedside, local, or remote?
Packaging: Single or multiple doses?
Fresh or frozen?
Based on your technologys
characteristics, next build a sales
forecast that incorporates
a rcgulatory stratcgy, with timing
and target content for each filing (e.g.,
it is not sufficient to write File an
IND; map out laboratory, clinical,
and other data that each filing must
contain to schedule what must be
gathered at each point)
a manulacturing/proccss
development and scale-up plan,
including a plan for transitioning to
commercial GMP scale-up
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