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Host Cell Expansion |
Cell Culture Production |
Purification |
Bulk Formulation |
Bulk Enzyme |
Final Product |
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In-house Capabilities
For over four years, BioMarin
has been operating its cGMP (current Good Manufacturing
Practices) manufacturing facility located in Novato,
California. The 70,000 square foot facility includes
25,000 square feet of dedicated cGMP process manufacturing,
quality control, and research and development laboratory
space. The facility is licensed by the U.S. Food
and Drug Administration and the European Commission
to manufacture Aldurazyme® (laronidase), an
enzyme replacement therapy (ERT) for the treatment
of mucopolysaccharidosis I (MPS I). Today, it is
also being used to produce Naglazyme® (galsulfase),
an ERT for the treatment of mucopolysaccharidosis
VI (MPS VI).
BioMarin has sufficient manufacturing capacity
to meet worldwide demands for both Aldurazyme
and Naglazyme.
In addition to the cGMP facility,
BioMarin continues to operate process development
and pilot scale laboratories to further evaluate
investigational product candidates.
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Manufacturing of Enzyme
Replacement Therapies:
Process Overview
Aldurazyme and Naglazyme are recombinant forms of
human enzymes, alpha-L-iduronidase and N-acetylgalactosamine
4-sulfatase, respectively. Both products are manufactured
using continuous perfusion cell culture —a
system that allows for the continual ‘harvest’
(removal) of the desired enzyme —and utilize
genetically modified Chinese hamster ovary (CHO)
cell lines. It is the harvested enzyme that ultimately
becomes the therapeutic product following extensive
purification.
The following is an overview of the manufacturing
process developed by the
research and development staff at BioMarin. The
success of this process is made possible through
the dedicated and coordinated efforts of five
primary groups: Process Development, Manufacturing
Sciences, Manufacturing, Quality Assurance, and
Quality Control.
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Raw materials are received, sampled, and stored
in a quarantine-controlled area. The samples are
tested against specifications and then, if specifications
are met, are issued a certification of analysis.
The raw materials are then moved to the released
materials controlled area where they are stored
until needed in the manufacturing process. |
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Frozen host cells that have been genetically engineered
to produce a specific
enzyme are aseptically added to a sterile flask
of growth medium. As the cells
multiply, they are transferred to a larger flask
to expand the volume and allow
room for additional cell growth. This process is
repeated until there is enough
volume at a specific cell density to inoculate (introduce
the cells into) a 110 liter stainless steel production-scale
bioreactor.
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| Stringent Environmental Controls |
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| Cell culture inoculation
and production occurs in class 100,000
clean rooms. The air in this room is
changed through HEPA filters 45 times
per hour and is approximately 40 times
cleaner than office air. The final purification
steps take place under even higher air
classifications (Class 10,000). An environmental
monitoring program is in place to ensure
microorganism and particulate levels
are kept within the specified levels. |
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Inside the 110 liter bioreactors, cells continue
to grow and multiply. The cells are continuously
stirred and air and specific nutrients are added
through sterile
connections to support cell growth. Dissolved oxygen,
temperature, pH and
nutrient levels are closely monitored and controlled
within precise limits to
enhance both cell growth and enzyme production.
As the cells mature, they begin to secrete the desired
enzyme that will later be vialed and administered
to patients. Before this can happen, however, the
enzyme must first be harvested and purified—removed
from the growth medium and separated from the other
by-product proteins and molecules—and then
undergo final bulk formulation.
To separate the desired enzyme from other proteins
and the growth medium,
the cell culture fluid is passed sequentially
through multiple chromatography
columns that utilize differential modes of selectivity.
With each step, more
impurities are removed, resulting in higher specific
activity of the desired
enzyme.
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| High Efficiency Production |
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| Optimizing each step of the manufacturing process is an essential component in maintaining an economically viable process. |
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To separate the desired enzyme from other proteins
and the growth medium, the cell culture fluid is
passed sequentially through multiple chromatography
columns that utilize differential modes of selectivity.
With each step, more impurities are removed, resulting
in higher specific activity of the desired enzyme.
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An Ultra Filtration/Diafiltration (UF/DF) process
is used to formulate the enzyme into a solution
that can be safely injected into humans. During
UF/DF, the solution is first concentrated and then
exchanged with formulation buffer to
bring the product and the excipients (other controlled
ingredients that enhance
the shelf-life stability and improve the product's
functional characteristics) to the correct concentrations.
Once this is complete, the product is at the exact
concentration and composition that will be administered
to patients.
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The bulk formulation solution is shipped off-site
under validated conditions to a
specialized fill/ finish vendor where it is vialed,
capped and labeled under sterile conditions. The
labeled vials are stored in quarantine until examined
by
BioMarin’s Quality Assurance personnel. The
vials are sampled and tested for a
number of characteristics to ensure safety, purity,
potency, identity, quality and composition. When
test results indicate that the final specifications
have been met, Quality Assurance releases the vials
for distribution and use in patients.
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