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Viruses can be lipid-coated or non- enveloped. Virus inactivation works by one of the following two mechanisms:  By attacking the viral envelope or capsid.

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Presentation on theme: "Viruses can be lipid-coated or non- enveloped. Virus inactivation works by one of the following two mechanisms:  By attacking the viral envelope or capsid."— Presentation transcript:

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2 Viruses can be lipid-coated or non- enveloped. 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. Virus inactivation involves dismantling a virus’s ability to infect cells without actually eliminating the virus. FDA specifies that a clearance of at least 3 logs must be achieved for mAb production.

3 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

4 Virus Inactivation Tank Ion Exchange Chromatography Storage Tank and Diafiltration

5 StreamMass Flow Rate (kg/batch) ComponentMass Composition (%) Connection S Acetic acid Impurities mAb WFI From storage tank S Polysorbate 80 WFI Chemical inlet S Nitrogen Oxygen Vent to atmosphere S Acetic acid Impurities mAb Polysorbate 80 WFI To polishing filter

6  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

7 SuperPro® Specification for Polysorbate80 at 0.01% wt Value Volume L Max Allowable Working Volume 90% Height/Diameter Ratio 3 Height 2.138m Diameter 0.713m Design Pressure1.52bar

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

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

10  Develop a way to simulate active and inactive virus concentrations › Optimize for temperature, holding time, and S/D concentration  Polysorbate 80 concentration of wt%  Purchase a used blending tank from Aaron Equipment Company for $8000

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