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