Transcript Viral Inactivation Unit

Virus inactivation involves dismantling a virus’s ability to infect
cells without actually eliminating the virus.
Viruses can be lipid-coated or nonenveloped.


Virus inactivation works by one of the
following two mechanisms:
By attacking the viral envelope or
capsid and destroying its ability to
infect or interact with cells.
By disrupting the viral DNA or RNA and
preventing replication.
FDA specifies that a clearance of at least 3 logs must be
achieved for mAb production.
Solvent/Detergent (S/D) Inactivation
• Effective with lipid-coated viruses
• Disrupts the interactions between molecules in the lipid coat ,
rendering the coat dysfunctional and impeding replication
• Commonly used and has a reliable safety record
• Chemicals need to be removed downstream
Pasteurization/Heat
• Effective with non-lipid and lipid-coated viruses
• Typically in a liquid at 60oC for 10 hours
• Protein product must have a higher thermal resistance than the virus
• Requires the addition of stabilizers, which will need to be removed
downstream
Low pH Treatment
• Most effective with lipid-coated viruses
• Acidic conditions deactivate virus
• Protein product must have a higher resistance to low pH conditions
than that of the viral particles
• No additional clearance steps are needed downstream
Virus Inactivation Tank
Storage Tank and Diafiltration
Ion Exchange Chromatography
Stream
Mass Flow Rate
(kg/batch)
S-042
763.2967
S-045
S-110
0.153
0.905
S-046
763.449
Component
Acetic acid
Impurities
mAb
WFI
Polysorbate 80
WFI
Nitrogen
Oxygen
Acetic acid
Impurities
mAb
Polysorbate 80
WFI
Mass Composition
(%)
0.0784
0.1064
3.2681
96.5470
50.0000
50.0000
76.7118
23.2882
0.0784
0.1064
3.2675
0.0100
96.5377
Connection
From storage tank
Chemical inlet
Vent to atmosphere
To polishing filter




There was no way to measure concentrations of active and inactive viruses
Manipulating temperature, holding time, and flow rates had very little effect
in our simulated unit
Theoretically, a longer holding time and a higher S/D concentration should
results in higher levels of inactivation
The volume increased minimally as the detergent concentration increased
SuperPro® Specification for
Polysorbate80 at 0.01% wt
Volume
Max Allowable
Working Volume
Height/Diameter Ratio
Height
Diameter
Design Pressure
Value
852.88 L
90 %
3
2.138 m
0.713 m
1.52 bar
Used
Supplier
Range of Cost
For Tank with agitator
Machinery and Equipment
Company, Inc.
$12,500.00
Aaron Equipment Company
$8,500.00 - $10,000.00
New
Cost for
Cost for Tank
Mixing Unit
without
agitator
Total Cost of
Tank and Agitator
$25,000.00 - $100,000.00
Slimline Manufacturing
$1,000.00
Pope Scientific Inc.
$11,000.00
Apache Stainless Equipment
Corporation
$15,000.00
$127,500.00
$128,500.00
$20,000.00
$35,000.00
Simulation Unit Operation:
In Place Steaming  Transfer In  Pull In  Holding  Transfer Out  In Place Cleaning
Operating conditions

Steam: normal pressure, 42.5 kg/batch

Polysorbate 80: 0.0765 kg/batch
Operating Costs
Cost
Steam (per batch)
$0.18
Polysorbate 80 (per batch)
$2.32
Usage Rate (per equipment-hr)
$2,088.00
Labour Hours (labour hrs/hr)
8
TOTAL $2,090.50 + 8 Labour Hours
Develop a way to simulate active and
inactive virus concentrations
› Optimize for temperature, holding time,
and S/D concentration
 Polysorbate 80 concentration of 0.01-1.0 wt%
 Purchase a used blending tank from Aaron
Equipment Company for $8000
