Dock to Stock at Skyline Biotech
A brief Outline of how we make our
product at Skyline Biotech
What is a Tech expected to do?
• Focus on
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Quality
Your training
The facilities around you
The Utilities that you use
• Operational Excellence
Dock to STock
• A process overview
• How exactly do we go from our raw
materials to a finished product
Alpha Amylase
From Bacillus subtilis
Training Goals
• Describe the main functions of the
manufacturing process
• Explain how these processes come together
to make a product
Master cell Bank
• Large volume of the original Strain or
construct is held under cryogenic
preservation.
– Usually in 1ml ampoules
– Usually in liquid Nitrogen
• Access is limited to ensure integrity of
product.
Working Cell bank
• Created from the
master cell bank for
producing batches of
product
• Do not need to
continually go to
Master cell bank
• A culture of the strain
that makes your
product
– Continuously checked
for efficiency and
purity
– When parameters are
not met then another
WCB is made from
MCB
Raw Materials
• Media for the cells
• A list of accepted
• Quarantined and tested
suppliers is kept
for quality before
• A list of accepted
being used in
substitutes is kept
manufacturing
• Many positions exist
to ensure the
consistency and
quality of the Raw
Materials chain
Process Paperwork
• Forms, Formulas and
SOP are kept current
and up to date
• Small changes may be
made but must be
proved to not change
the process
• All completed
paperwork is kept
• Typically people with
experience in writing
or the process are
involved
Small Scale Cell culture
• A sample from the
WCB is transferred to
successively larger
culture tubes
• Selective Media used
to keep strain
• Check for
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Cell density
pH
Temperature
Nutrient concentration
• Aa, sugar, oxygen,
carbon dioxide
Continuously check for
• Quality
• Quantity
• consistency
Large cell Culture
• Typically a culture is
not diluted more than
10x between each
fermentation exchange
• Fermentation process
is Batch
– Cells are grown until
they reach a certain
density then transferred
en mass to a new
vessel with new media
How important is sterile
technique?
Product Recovery
• Harvest. This depends on where the
product is
– In cells
• Separate the cells out, break open cells and start
product purification
– In media
• Separate the cells out, start product purification from
the spent media
Types of Harvest
• TFF Tangential Flow Filtration
• Continuous Flow Centrifugation
Purification, Isolation and Viral
inactivation
• TFF or column chromatography is used to
separate proteins by size, charge, affinity or
hydrophobic.
• The product will be separated by a complex
mixture
• The product may not be absolutely pure
– However if the process is the same the contaminants are
validated with the final product
Viral Inactivation
Formulation and pool bulks
• Formulation
– Concentrated protein is formulated to become
tolerable to the patients
– Ensures that the product is compatible in blood
pH 7.1
– Bulk is frozen and stored in freeze/thaw tanks
to await filling
Filling
• Final vials are filled under as sterile
conditions as possible
Freeze drying
• Process increases shelf life and product
stability
Product inspection
• Empty vials and stoppers are checked
• Filled are checked
• Also perform a container integrity test
Label Final Product
Final Product is packaged.
Alpha Amylase is ready to use