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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– 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• Quarantined and tested
for quality before being used in manufacturing
• Many positions exist to ensure the consistency and quality of the Raw Materials chain
• A list of accepted suppliers is kept
• A list of accepted substitutes is kept
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– 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