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

3

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

Cen

trifu

gal f

orce

Flui

d flo

w fo

rce

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+++

+++

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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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Physical Changes kSep

Pinch valve splash cover / drip trayPressure Sensors

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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

Thank you