Virus inactivation involves dismantling a virus’s ability to infect cells without actually eliminating the virus.
Viruses can be lipid-coated or nonenveloped.
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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 60 o C 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
S-042
S-045
S-110
S-046
Mass Flow Rate
(kg/batch)
763.2967
0.153
0.905
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

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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

Supplier
Used
Range of Cost
For Tank with agitator
Machinery and Equipment
Company, Inc.
$12,500.00
Aaron Equipment Company $8,500.00 - $10,000.00
Cost for
Mixing Unit
New
Cost for Tank without agitator
Total Cost of
Tank and Agitator
$25,000.00 - $100,000.00
$128,500.00
Slimline Manufacturing
Pope Scientific Inc.
$1,000.00
$127,500.00
$11,000.00
Apache Stainless Equipment
Corporation
$15,000.00
$20,000.00
$35,000.00

Simulation Unit Operation:
In Place Steaming  Transfer In  Pull In  Holding  Transfer Out  In Place Cleaning
Operating conditions
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Steam: normal pressure, 42.5 kg/batch
Polysorbate 80: 0.0765 kg/batch
Operating Costs
Steam (per batch)
Polysorbate 80 (per batch)
Usage Rate (per equipment-hr)
Labour Hours (labour hrs/hr)
Cost
$0.18
$2.32
$2,088.00
8
TOTAL $2,090.50 + 8 Labour Hours

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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