Risk Group 2 Biosafety Containment Challenges Surrounding Single-Use Fluidized Bed (kSep) Centrifuge Technology
Francine Preston* (IBC Chair) and Ravi Bhatia (Cell Technologies)Janssen R&D, Spring House, PA
2014 CDC Biosafety Symposium Conference
*Presenter
Development Project
Scale up <1000L of RG-2 Agent Evaluate kSep technology to harvest cells
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Single-Use Centrifuge - KSep
Bioreactor
Fluidized Bed Centrifuge (kSep Systems)
Single-Use System (All product contact components are disposable)
Fully automated system
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HMI
Tubing holders
PumpsRotor & chamber
Single-Use Chambers
Rotor
Principle of kSep®
• Counteraction of Centrifugal force and fluid flow force• Creates fluidized bed of cells• Cells remain in suspension• Very low shear
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Flui
d flo
w fo
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Horizontal axis of rotation
Risk Assessment
Biohazard Agent Registration (BAR) reviewed by IBC. BSL2 Large Scale Containment. Known infectious agent transmitted via mucous
membrane and aerosol route, effective treatment available.
Prevent potential exposure to employees and environment– Engineering and Administrative Controls and PPE
Medical Surveillance Emergency Response Plan FMEA to further evaluate kSep containment
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IBC Approval Process
Approval Granted in 3 Phases Phase 1 – RG-2 materials at small scale reactor Phase 2 – Utilize kSep to harvest RG-2 cells
– Phase 1 Controls plus – kSep FMEA
Phase 3 – Scale-up process to < 1000L – Phase 2 Controls plus – Modify facility controls to meet BSL2 LS requirements
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Phase I –
Small Scale Reactor Control Access Contain Process
– Biosafety Cabinet – Reactor Closed System with HEPA (equivalent)
Vented filter Medical Surveillance Personal Protective Equipment Emergency Response Signs and Labels on process
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Medical Surveillance
Immunization required Immune Compromised must be evaluated by
medical Workers with Acute respiratory infections are
discouraged from working on the process until infections subsides
Post Exposure Prophylaxis provided
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Personal Protective Equipment
kSep operation / Access closed system Disposable Coverall Double nitrile gloves Double booties Hairnet Powered Air Purifying
Respirator (comfort)
Room Access / Reactor Observation
Disposable Coverall Double nitrile gloves Double booties Hairnet Safety glasses Face mask
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Emergency Response
Spill Response Small spills cleaned by operator Large spill cleaned by outside company
Employee Exposure Evaluated by Occupational Health Post Exposure Prophylaxis will be considered
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Phase 2 –
kSep FMEAEvaluate equipment and facility controls Controls for Disposable Bioreactor established.
kSep technology new to RG2 infectious agents– Loss of containment. Any exposure will far exceed
infectious dose.– Implement controls for all identified risks.
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kSep
Physical changes– Splash guard over pinch valve panel on the side of
machine – Implementation of pressure sensors at 3 locations– Change tubing size
Programing changes to allow early leak detection prior to introducing cell suspension – priming system
Procedural changes to add additional level of protection against release– Operator present– Frequent Equipment Preventative Maintenance
Emergency Stop button installed near exit
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Phase 3 –
BSL 2 Large Scale - Facility Controls Airflow
– Negative to surrounding areas– Single – pass HEPA filtered
Cell Culture Room – Floors Epoxy Resin (No Open
floor drains)– Berm in floor to prevent flow
into airlock– Floor-Wall junction protected
by several inches of Epoxy coating
– Latex-painted gypsum wallboard walls
– Sealed tile ceilings
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• Airlock– Negative to corridor– Gowning – Spill Supplies
• Secondary Containment– Pallets Beneath
Reactor – Floor Berm