Gene Therapy Production Facility Considerations Robert Sausville Center for Biologics Evaluation and Research Office of Compliance and Biologics Quality Division of Case Management 11/15/2018

Overview General Considerations Special Considerations Biosafety Levels Air Quality Environmental Monitoring Validation Qualification Miscellaneous

Overview New manufacturing areas retrofits of existing facilities located within existing facilities Designed to minimize potential for contamination between products between early and late stages of production Operations should be well controlled Personnel should be appropriately trained someone responsible for facility operations, equipment validation and maintenance, record keeping

General Considerations Good techniques and appropriate equipment used to minimize exposure to infectious agents Strict adherence to standard practices and techniques (consistent manufacture of product) Safety equipment (primary barrier) biological safety cabinet closed containers other designs to minimize aerosols Facility design (secondary barrier) amount of protection depends on the nature of the infectious agent and production associated risks (aerosols)

General Considerations cont. As the risk for aerosol transmission increases, higher levels of primary containment and multiple secondary barriers may become necessary

General Considerations cont. Facility designed for aseptic processing Smooth surfaces, seamless tile, etc. Hard, easily cleanable and impervious finishes Non shedding ceilings HEPA filtered air from seperate air handling unit separation of production from other areas of the facility (often a hospital) Phase 1/II Class 100,000: general manufacturing areas (10,000 III) Class 100: all open manipulations

General Considerations cont. Separate entrance/gowning area Air locks (containment) No personnel access between vector production and transduction areas Room(s) for support areas buffer media prep, glass/equipment wash Storage areas raw materials, cell banks adjacent to production if possible to minimize traffic in and out of the facility

General Considerations cont. Material and personnel flows designed to maximize efficiency and minimize mix-ups unidirectional flows where possible also controlled by procedures or temporally Emergency power for critical systems (UPS)

General Considerations; Control of the Facility Production area(s) should be separate from other activities (research, testing) Potential cross-contamination should be minimized between process steps, and other production activities Access into the production area should be limited Equipment used in production should not be shared with non-production activities Environmental monitoring and product testing should be performed to ensure adequate control of the process/area Spill/accident procedures should be in place

Special Considerations; Multi-use scenarios Concurrent vs. campaign production will impact design considerations Personnel work on one cell line at a time Procedures in place to prevent cross contamination Appropriate changeover procedures Adequate segregation of concurrent activities color coded labeling bar coded The sponsor is responsible for assuring that the contract manufacturer has all the procedures in place

Special Considerations Cont. Potential Routes of Cross Contamination Centrifuges (generation of aerosols) one sample processed at a time cleaning/sanitization of centrifuges between “lots” Pipettors effective cleaning procedures filters Incubators

Appropriate BioSafety Level In general, BioSafety Level 2 Transduction Higher risk, BioSafety Level 3 Vector Preparation Defined in CDC-NIH publication Biosafety in Microbiological and Biomedical Laboratories

BioSafety Level 2 Transduction and non-viral vector preparation access limited personnel trained in handling pathogenic agents infectious wastes are decontaminated before disposal gowning required (lab coat, hair cover) gloves should also be worn for aseptic manipulations Class I or II Biosafety Cabinets to be used: for procedures potentially creating aerosols aerosol generation should be minimized with high concentrations or large volumes of infectious agents

BioSafety Level 3 Viral Vector prep areas, higher levels of containment Negative pressure or “sink” for containment All activities with infectious materials are conducted in biological safety cabinets Class I, II or III may be used Passage between two sets of doors is a basic requirement An autoclave for decontaminating waste is available preferably in the laboratory Ducted exhaust provided not recirculated may be discharged to the outside without being filtered

BioSafety Level 4 Class III biosafety cabinets or personnel in suits with life support systems All materials must be autoclaved before leaving BSL4 area Exhaust air HEPA filtered

Air Quality Recommend that production areas receive single pass air (no recirculation) for vector production Dedicated air handler where possible Segregating air supply from rest of facility important, (hospital setting) HEPA-filtered air Objective: to meet Class 100,000 specifications for Phase I/II In-line vs. terminal

Air Quality cont. Air pressure differentials between areas should be balanced to maintain cleanliness or containment Positive (aseptic processing) Negative (containment) for steps needing greater than BSL-2 Open steps in biosafety cabinets (Class II) Quality monitored to assure facility is acceptable for production

Environmental Monitoring Testing of surfaces and for viable and non-viable particulates Demonstrate facility under control as part of validation routine monitoring program Frequency and intensity dependent on how close to GMPs the facility will operate specifications based on desired Class

Environmental Monitoring cont. Viable particulates active air monitoring devices (slit to agar, centrifugal samplers) settling plates (passive, less desirable) Non-viable particulates particle counters room classifications, certification of biosafety cabinets Surface “contact” plates or swabs monitor cleaning efficacy and personnel asepsis

NASA Standards; Viable Air Particulates Room classification, defined by Federal Standard 209E, determined by non-viable particulate monitoring under dynamic conditions. Class 100,000 2.5 CFU/ ft3 Class 10,000 0.5 CFU/ ft3 Class 100 0.1 CFU/ ft3

Settling Plates Exposure times Class 100,000 1 CFU/ 9cm plate 0.11 hours 2 CFU/ 9cm plate 0.21 hours 1 CFU/ 14cm plate 0.04 hours 2 CFU/ 14 cm plate 0.09 hours Class 100 1 CFU/ 9cm plate 2.65 hours 2 CFU/ 9 cm plate 5.31 hours 1 CFU/ 14 cm plate 1.10 hours 2 CFU/ 14 cm plate 2.21 hours

Validation “Establishing documented evidence which provides a high degree of assurance that a specific process will consistently produce a product meeting its predetermined specifications and quality attributes.”

Validation “Sliding Scale” approach for clinical manufacturing Facilities involved in clinical manufacturing should be in compliance with the concepts of cGMPs Do not expect full validation in early stages (may not have 3 repetitive runs, worst case configuration, etc.) facility supplying clinical material to other institutions implies that it meets cGMPs and should approach what is expected for commercial facilities Phase 3 material should be manufactured at close to full cGMP

“Sliding Scale” Approach, An Example Autoclave used to prepare sterile materials Early (I/II) demonstrate proper cycle achieved monitor temperature, pressure, and time use of biological indicators for verification loads not well defined

Autoclave cont. Middle (II/III) temperature mapping done to determine cold/hot spots biological indicators placed to verify cycle at problems points loads are somewhat defined

Autoclave cont. Late (III+) lethality of cycle determined at monitored points loads are well defined and standardized each load configuration has been mapped or worst-case load has been validated Another example Sanitizer effectiveness Phase I/II supported by literature Phase III supported by validation

Process Validation Sterilization Decontamination Aseptic Processing Cleaning Inactivation/removal of adventitious agents and other contaminants

Equipment Qualification Program in place to demonstrate that equipment operates as expected Should include periodic monitoring

Equipment cont. Air handlers Biological safety cabinets pressures filter integrity airflows; velocities leak testing

Equipment cont. Incubators uniform temperature carbon dioxide filters Centrifuges speed vacuum (ultras) temperature

Equipment cont. Autoclaves temperature pressure kill cycle Lyophilizers shelf temperature vacuum

Raw Materials Critical raw materials - established criteria for acceptance from vendors: sterility adventitious agents activity/purity Vendor’s Certificate of Analysis identity test where possible Inventory Control proper storage FIFO

Water Should meet Water for Injection (WFI) specifications : microbial <10 CFU/100ml endotoxin <0.25 EU/ml chemical tests per USP WFI for all product contact surfaces and formulations May be purchased

Personnel Designated person in charge of facility Responsible for: limiting access training maintenance/safe operations writing and enforcing procedures Production personnel are trained (periodic retraining) Appropriately gowned for production step

Final Thoughts Design facility for worst-case = maximum flexibility Consider filing a Master File for facilities handling several IND products Meet with FDA to discuss your Phase I/II (or III!) facility