Mammalian Cell Line Engineering and Biopharmaceutical Manufacturing

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Mammalian Cell Line Engineering and
Biopharmaceutical Manufacturing
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Flexible biopharmaceutical product development
Catalents flexible business model allows
customization to fit your requirements. Whether
you need our proprietary GPEx mammalian cell
engineering technology or contract services to
work with your cell line, we can provide
virtually any combination of the following
services under a single project manager
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Why GPEx technology?
Faster
Production of a master cell bank candidate from
clients CDNA in 5 months or less
More reliable
Consistent expression stability and high
production levels with no need for selection,
ever!
Choice of mammalian cell lines (class switching
and chimeric antibody production), co-expression
of different genes in one cell line...with fast
timelines
Flexible
Proven
Established worldwide regulatory path
Good value compared to competitors and favorable
intellectual property
Economic
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GPEx technology is faster

• Using conventional gene insertion techniques, it
  can take 18 months or longer to produce a master
  cell bank candidate. Our GPEx technology reduces
  this time to 5 months or less.
• Unlike conventional methods, GPEx transductions
  are stable and occur in virtually 100 of target
  cells. Because of this high level of efficiency,
  there is no need for antibiotic selection,
  shortening the master cell bank production time
  significantly.

2 weeks
2 weeks
2 weeks
2 to 3 months
Clonal cell line selection and expansion
Single subunit protein
Master cell bank candidate
Process development and cGMP manufacturing
Retrovector transduction of cells
Limited dilution cell cloning
DNA cloning
Monoclonal antibody
4 to 5 weeks
4.5 to 5 months
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GPEx technology is faster (contd)

• The stable cell pools produce small amounts (mg
  to g) of variant antibodies prior to the
  selection of a high-expressing clone. This allows
  for screening of variants and optimal selection
  of final clinical candidates. The target protein
  and its variants can be provided in as little as
  2.5 months for analysis and characterization.
  The pools then become the starting cells for MCB
  candidate selection, greatly reducing timelines.

2 weeks
2 weeks
2 weeks
4 weeks
DNA cloning and sequence verification
Target protein for analysis and characterization
Retrovector production
Pooled cell line
2.5 months
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GPEx technology is more reliable
Consistently higher expression levels
Reliability
GPEx technology consistently produces cell lines
with higher expression levels than other
technologies. The gene insertion process
specifically targets the active regions of the
host cell genome (it is not a random process). In
addition, GPEx employs an iterative insertion
process that drives up the inserted gene copy
number and proportionally increases protein
expression levels. This leads to higher desired
protein yieldstypically 30 to 100
picograms/cell/day.  
Antibody Titer (g/L)
Specific Productivity (P/C/D)
High Titers (over 2 to 4 g/L)
High Productivity (over 30 to 100 p/c/d)
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GPEx technology is more reliable (contd)
High Stability Reliability
GPEx technology inserts are more stable than
competing high copy number systems because each
insertion event takes place at a different
genomic location. This prevents the occurrence
of recombination events that lead to DNA loss
and/or DNA rearrangement.
Real-time PCR analysis
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GPEx technology is more reliable (contd)
The high transduction efficiency, coupled with no
antibiotic selection requirement, allows easy
titration of heavy chain (HC) and light chain
(LC) genes to the correct gene ratio to yield
maximum antibody production and cell line
stability. The optimum HC/LC ratio remains
constant from generation 0 to 60 due to the HC
RNA and LC RNA individual stabilities.
Real-time PCR analysis
Real-time PCR analysis
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GPEx technology is more reliable (contd)
Reproducibility Reliability
Commercial mAb X
Commercial mAb X produced with GPEx technology
both antibodies lead to
Indistinguishable tryptic map chromatograms Equi
valent bioactivity Identical glycoforms Levels
of individual glycoforms within the commercial
specifications
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GPEx technology is flexible

• GPEx technology can be applied to a wide range
  of cell types, including all mammalian cells.
• Backbone vectors containing the gamma constant
  region for human IgG1, IgG2, IgG3, or IgG4 and
  light chain kappa or lambda constant regions
  allow fusion of DNA sequence encoding IgG
  variable regions to any constant regions in the
  GPEx Retrovector. These constant regions can be
  mixed and matched to produce any IgG molecule
  desired.
• Co-expression of multiple, different genes is
  advantageous for mAb production and applications
  that require production of 2 or more recombinant
  proteins in a cell line. Because of GPEx high
  efficiency in the insertion process, the need for
  multiple selectable markers is avoided. In
  addition, the levels of each gene can be
  titrated (varying the ratio) to yield the desired
  result.

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GPEx technology has high potential
Very high specific activity (P/C/D) Flexibility
The high specific activity of the GPEx cell
lines allows additional upstream process
development to greatly increase titers of the
target protein.
Antibody U upstream process development
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GPEx technology is proven

• Over 125 different mAb and mAb fusions and over
  25 different recombinant proteins have been
  produced using the GPEx system.
• Clinical trial status
• Proteins expressed by GPEx cell lines are in
  clinical trials in Asia, Australia, and Europe
  clinical supplies produced either at
  Catalent-Middleton or clients facility
• Antibody manufactured by client using a GPEx
  cell line has been approved and marketed in Asia
• The cGMP production of an antibody for use in a
  clinical trial in South America is in progress
  at Catalent-Middleton
• Client has initiated a US-based Phase I clinical
  trial (patients dosed) using material expressed
  by a GPEx cell line that has been manufactured
  at Catalent-Middleton facility
• Second US IND approved for an antibody produced
  by a GPEx cell line

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GPEx technology economic advantages

• GPEx technology is a good value and short
  timelines allow you to turn your product into
  revenue faster.
• Intellectual Property
• GPEx technology is patent protected
• GPEx technology is well positioned within the
  current antibody patent space and avoids many of
  the mammalian cell engineering patents

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GPExâ technology conclusions
GPEx is the first step to bring your protein
from discovery to manufacture and into clinical
trials.

• Advantages of GPEx technology
• High speedMaster cell bank candidate cell lines
  are produced in 5 months. Protein is available
  for analysis early in the cell line development
  timeline.
• ReliabilityThrough consistently producing highly
  stable, high expressing/high yielding cell lines.

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GPExâ conclusions (contd)

• FlexibilityWide range of cell types, including
  all mammalian cells, can be used
• Antibody class switching.
• Co-expression of different genes in one cell
  line.
• Proven recordOver 100 different mAb and mAb
  fusions and over 25 different recombinant
  proteins have been produced with GPEx. FDA IND
  approval.
• EconomicWell priced in comparison to competitors.

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Welcome to our Catalent Middleton site

• 43,000 square feet
• cGMP manufacturing and QC laboratories
• RD molecular and cell biology labs
• RD process development and analytical
  laboratories
• Equipped to develop and manufacture small-scale
  antibody and recombinant clinical products from
  mammalian cells

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Middleton site capabilities

• Molecular and cell biology
• Vector design/optimization
• Cell line development
• Cell banking
• Cell culture process development
• Up to 100L scale
• Downstream process development
• Analytical support and stability studies
• Preformulation/formulation development
• Clinical manufacturing (up to 200L scale)
• Technical transfer to commercial sites

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Contact us
We have outstanding technology, a state-of-the
art facility, and a knowledgeable and experienced
team. For more information about working with
GPEx technology, or to learn more about our
manufacturing services, call us at [phone]
or email us at gpexinfo_at_catalent.com. Take that
first step to bring your product to market
faster contact us!
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