FREEZE DRYING OF

BIOPHARMACEUTICALS
FREEZE DRYING
• Process takes place at low temperatures, which in general
minimizes the in-process deterioration, resulting in dried
products with improved stability and ease of handling as
compared to the products in liquid state

• Most common application of freeze-drying involves the

manufacture of injectable products & in some cases, it is also
used to generate oral solid products with fast-dissolution
property
OBJECTIVES
• To preserve the biological activity of a product.

• To reduce the product weight to lower the transportation cost.

• To extend the shelf life or stability.

• To dry thermolabile materials.

• To eliminate the need for refrigerated storage.

• To get accurate, sterile dosing into the final product container.

 STAGES INVOLVED
LOADING

FREEZING

PRIMARY DRYING

SECONDARY DRYING

SEALING
1. LOADING

• The freeze-drying process for pharmaceutical injectable

products starts with filling the liquid or semiliquid product
aseptically into vials, which are then partially stoppered
with elastomeric closures (stoppers)
2. FREEZING

• Freezing is generally the first step in a freeze-drying process,

in which nearly 90% of the water is converted to ice crystal

• All solutes in the formulation are solidified into a matrix

either in amorphous or crystalline state, or in a mixture

• The conversion from water to ice crystals starts with ice

nucleation, which is followed by ice crystal growth
• Ice nucleation temperature is often in the range of 10–15°C
or more below the equilibrium freezing point : SUPER
COOLING

• Freezing is a very critical stage in a freeze-drying process

• Annealing : Facilitate the crystallization of bulking agent

• After annealing, the product temperature is generally

lowered to a final temperature and held long enough to
complete solidification
3. PRIMARY DRYING

• The ice crystals formed in the freezing stage are removed by

sublimation in the primary drying stage

• Performed at relative low temperature and low pressure, and

the solute structure formed during freezing will be retained

• It is performed safely below the product’s critical collapse

temperature
• Chamber pressure & shelf temperature are interrelated, and
integrally determine the ice sublimation rate

• In most cases, the optimized chamber pressure in primary

drying is in the range 50–200 mTorr

• Primary drying is normally the most time-consuming stage

SECONDARY DRYING
• Removal of unfrozen water

• Chamber pressure will be the same

• Shelf temperature for the secondary drying should be high,

normally between 30◦C and 50◦C.

• Continued until the product reaches acceptable moisture

content for long term storage
• Depending on the application, moisture content in fully dried
products is typically between 0.5% and 3%

• In most cases, the more dry the product, the longer its shelf life
will be

• During secondary drying, a “sample thief” mechanism may be

used to periodically remove vials from the freeze dryer for
residual moisture content determination.
CYCLE OPTIMIZATION
1. Freezing and annealing

2. Thickness of the product

3. Critical Collapse Temperature

SEALING
• Lyophilized products are extremely hydroscopic and they
must be sealed in air tight containers to prevent rehydration
from atmospheric exposure

• Freeze dryers can be configured with a “stoppering”

capability to seal the product while it is still under partial
vacuum inside the unit
ADVANTAGES
• Removal of water at low temperature

• Thermolabile materials can be dried.

• Compatible with aseptic operations

• More precise fill weight control

• Sterility can be maintained

DISADVANTAGES
• Many biological molecules are damaged by the stress
associated with freezing, freeze-drying, or both.

• The product is prone to oxidation, due to high porosity and

large surface area. Therefore the product should be packed in
vacuum or using inert gas or in a container impervious to
gases

• Cost may be an issue, depending on the product

• Long time process

REFERENCES
• Michael C. Flickinger (2013) Downstream Industrial
Biotechnology. John Wiley & Sons, Inc.