Single Use Bioreactor Scale up: Introduction of a 2000L SUB Equipment Train in 12 months

Single Use Bioreactor Scale up: Introduction of a 2000L SUB Equipment Train in 12 months Finesse CellWorld San Fransisco Claire Walsh BSc, MSc BioProcess Skills and Technology, Operations, Janssen Sciences Ireland ONE TEAM Making the Difference for Patients WORLDWIDE 0 Finesse CellWorld San Francisco

Presentation overview: My Story Janssen Sciences Ireland Janssen Process overview Brief overview of the original 1000L SUB project Project drivers and scope Timeline Outline of the 2000L SUB Project Project Drivers and scope Timelines Engineering Run data, including Raman probe data Benefits and Limitations Q & A Finesse CellWorld San Francisco 1

My Story Completed a BSc in Biochemistry and an MSc in Biotechnology in University College Cork Completed a Student Placement in PDMS in Janssen Travelled and worked in Australia before returning to Janssen to work in multiple roles including New Process Development Projects, Global Standardisation Projects, New Product Introductions/Tech transfers, Daily Process Support and Engineering Projects Finesse CellWorld San Francisco 2

Cork Janssen Pharmaceutical Sciences Janssen Sciences Ireland DePuy Janssen R&D Limerick Vistakon Dublin J&J Consumer J&J Medical Janssen Alzheimer Immunotherapy Janssen (Marketing) Finesse CellWorld San Francisco 3

Cork Janssen Pharmaceutical Sciences Janssen Sciences Ireland DePuy Janssen R&D Limerick Vistakon Dublin J&J Consumer J&J Medical Janssen Alzheimer Immunotherapy Janssen (Marketing) Finesse CellWorld San Francisco 4

Janssen Sciences Ireland Key Products include: Simponi, Stelara, Darazalex, Prezista, Invega Sustenna and Risperidal Finesse CellWorld San Francisco 5

Focus for the future Finesse CellWorld San Francisco 6

Focus for the future Finesse CellWorld San Francisco 7

Janssen Process overview Commercial Suite Stainless Steel Bioreactors Perfusion process Clinical Suite (SUBs) Single Use Technology Fed batch process Finesse CellWorld San Francisco 8

Janssen Process overview Commercial Suite Stainless Steel Bioreactors Perfusion process Clinical Suite (SUBs) Single Use Technology Fed batch process Finesse CellWorld San Francisco 9

Janssen Process overview Commercial Suite Stainless Steel Bioreactors Perfusion process Clinical Suite (SUBs) Single Use Technology Fed batch process Finesse CellWorld San Francisco 10

Janssen Process overview Commercial Suite Stainless Steel Bioreactors Perfusion process Clinical Suite (SUBs) Single Use Technology Fed batch process Finesse CellWorld San Francisco 11

Disposable Cell Culture Process Train: Project Drivers Decision to move JNJ Platform from Perfusion to Fed Batch Historically Janssen Cell Culture Processes were perfusion based Outsource of Upstream Fed Batch Processing to CMOs Limited in house experience in Fed Batch Decision made to provide back-up internal supply chain capacity for Fed Batch material Finesse CellWorld San Francisco 12

1000L SUB Project Scope Capability for pre-clinical, clinical and commercial production Project must deliver 2 x 1,000 Single Use Bioreactors The capability to have 2 Single Use Bioreactors in operation with 9 months notice Project cannot interfere with the current clinical / commercial manufacturing schedule or approved Operational areas Project must cost less than $21m including all works The new suite must be integrated into existing Automated Systems Multi product capability on a campaign basis Maximize use of disposable technologies Finesse CellWorld San Francisco 13

Why choose to go disposable? Batch Turnaround time Product change over capabilities Cleaning validation requirements Disposable technology supports pre-clinical/early phase production Installation and Qualification Reduction in overall IOQ activities In-house historical disposables experience Finesse CellWorld San Francisco 14

What was the equipment selection criteria? The following criteria was applied to the equipment and disposables selection process: Bag film defined as CX514 material An Integrator with GMP experience Availability of novel disposables The control system needed to integrate into the existing site systems Customizable Assemblies Finesse CellWorld San Francisco 15

What did the Project Deliver? Fully Disposable PreCulture Train Single Use Bioreactors (SUBs) 1 x 250L Seed SUB 2 x 1000L Production SUB Single Use Mixers (SUMs) Harvest Collection SUM DPC (Chromatography) Steps Media Prep Disposable Bio-containers for sampling and media/buffer storage Extensive array of disposable assemblies Finesse CellWorld San Francisco 16

Project timelines October 2010 to July 2011 Engineering Design, Procurement and Construction (10 months) July 2011 to November 2011 Commissioning and Qualification and Engineering Runs (16 weeks) December 2011 to Feb 2012 First Phase 1 Clinical Campaign (12 weeks) Finesse CellWorld San Francisco 17

Original Suite Layout Finesse CellWorld San Francisco 18

Cell Culture Suite 2 Layout Finesse CellWorld San Francisco 19

What happened next? Huge amount of experience gained using disposables On-going Process Improvements building in extra reliability and decreasing both set-up and turnaround times Increasing demand in the clinical suite Requirement to increase capacity Finesse CellWorld San Francisco 20

2000L SUB Project Scope Additional fed batch capacity required Install and Qualify a 2000L SUB and associated equipment train Detailed design required in order to maximize capacity with footprint available Relocate an existing processing suite to remodel for 2000L SUB Suite handover within 12 months Project must cost less than 5.1m including all works Finesse CellWorld San Francisco 21

Cell Culture Suite 2 pre-2000l SUB Finesse CellWorld San Francisco 22

Cell Culture Suite 2 incl. 2000L SUB Finesse CellWorld San Francisco 23

Cell Culture Suite 2 Finesse CellWorld San Francisco 24

Cell Culture Suite 2 Finesse CellWorld San Francisco 25

Cell Culture Suite 2 Finesse CellWorld San Francisco 26

Cell Culture Suite 2 Finesse CellWorld San Francisco 27

Cell Culture Suite 2 Finesse CellWorld San Francisco 28

Cell Culture Suite 2 Finesse CellWorld San Francisco 29

Project timelines Oct 2015 to Mar 2016 CAR approval, Engineering Design, Procurement and Construction part 1 Mar 2016 to Aug 2016 Construction part 2 Equipment FAT Commissioning and Qualification Aug 2016 to Sep 2016 (12 weeks) Engineering Runs First GMP batch thawed in PreCulture 16 Nov 2016 Finesse CellWorld San Francisco 30

Complete Disposable Cell Culture Train PreCulture Fully Disposable 250L, 500L, 1000L and 2000L SUB Capacity 2000L and 3000L SUM Capacity DPC and VIN SUM capacity Finesse CellWorld San Francisco 31

Process Flow (up to Viral inactivation) Medi a 250L/500L Seed SUB to 1000L/2000L Production SUB 2000L/3000 L SUM Finesse CellWorld San Francisco 32

PreCulture Process Flow Historical experience with disposables technology here (common process to the Commercial suite) Flask Stages from 125mL to 500mL capacity Wavebag stages from 2L to 50L capacity Finesse CellWorld San Francisco 33

Seed and Production SUBs Seed SUB 250L and 500L Capacity DeltaV Recipe Driven Control Option to use as both a seed and a production SUB Process duration 4 days (Product dependent) Process parameters: ph, DO, Temp, Agitation Production SUB 1000L and 2000L capacity DeltaV Recipe Driven Control Process duration 21 days (Product dependent) Process parameters: ph, DO, Temp, Agitation Media and Antifoam addition Finesse CellWorld San Francisco 34

Harvest Train Centrifuge Use (not all products) Non-disposable Depth Filters (POD filters) Fully disposable assembly train Pressure sensors pre and post filter set-up Polishing filter post Depth Filters Harvest Collection SUM (2000L and 3000L capacity) Agitation Temperature control Sampling points Finesse CellWorld San Francisco 35

3000L SUM: New Technology Finesse CellWorld San Francisco 36

3000L SUM: New Technology Finesse CellWorld San Francisco 37

Process Monitoring in the SUBs ph Controlling probes: Electrochemical probe Optical ph probes in the process of being proved out DO Controlling probes: Electrochemical probe Optical ph probes in the process of being proved out Temperature Thermowell built into the disposable SUB bag Agitation Built in sheath in the disposable SUB bag Shaft inserted into the sheath prior to each batch This is then connected to the motor to drive the impeller Pressure Built in Pressure Sensor in the disposable SUB bag Finesse CellWorld San Francisco 38

Engineering Run Cell Culture Data Comparison 2000L SUB, 1000L SUB and Small Scale Satellite Run Raman Data Finesse CellWorld San Francisco 39

Cell Growth Comparisons Finesse CellWorld San Francisco 40

Cell Performance Comparisons Finesse CellWorld San Francisco 41

Metabolite Data Finesse CellWorld San Francisco 42

Raman Background Correlated the spectra to the off-line data Model developed for each key process parameter (eg: Glucose, Lactate) you re interested in measuring. Model development is time intensive Possible future applications include: Potential for use as an inline monitor, therefore reducing the reliance on off-line instruments Feed cycle optimisation * Research funded by the Irish Research Council Finesse CellWorld San Francisco 43

Raman Data: Glucose 2000L SUB 5L Scale Black Dots: Daily Off-line measurements Red Line: Raman Data Finesse CellWorld San Francisco 44

Raman Data: Ammonia 2000L SUB Black Dots: Daily Off-line measurements 5L Scale Red Line: Raman Data Finesse CellWorld San Francisco 45

Raman Data: Lactate 2000L SUB Black Dots: Daily Off-line measurements 5L Scale Red Line: Raman Data Finesse CellWorld San Francisco 46

Benefits System Set up time and turnaround time Increased Flexibility for Operations Cross contamination risk reduction C & Q: time savings Reduced Capital Investment and Rapid install time No Cleaning Validation requirements Ease of Product Change over Footprint saving Design Flexibility (Bag re-design, quick and easy) Incorporation of new technology (quicker) Great relationship and customer experience with Finesse Finesse CellWorld San Francisco 47

Limitations x Scale up limitations x Lack of a sight glass in the bag design x Lead times on bags x Leachables and Extractables studies x No confirmation of integrity pre-use x No pressure transfer x Minimum mixing volume (other options becoming available) Finesse CellWorld San Francisco 48

Acknowledgments Liz Dooley Nicola Sheehan Seán Coomey Anne O Leary Dan Olden 2K SUB Core Project Team Carl Rafferty (funded by the Irish Research Council) Engineering Team in Cork C & Q Team in Cork Operations team in Cork Automation Finesse and their Support Team Finesse CellWorld San Francisco 49

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Summary Implemented Fully Disposable Cell Culture Equipment Train Installed on time and within budget Minimal disruption to existing Operations Integration with existing site systems Replication of design including scale up Data to prove successful scale up In line with the Credo to customer and patients Finesse CellWorld San Francisco 51

Back-up Slides Finesse CellWorld San Francisco 52

Details sent to Cell World Business case for 2000L SUB train clinical capacity constraint etc. Background on the Design of the 2000L SUB Suite Engineering run and scale up data (including Ramen data) Comparison between Stainless Steel Manufacturing and Single Use Technology Abstract (max 200 words): 1. CREATIVE DESIGN Overview of the design process for the delivery of the new Cell Culture manufacturing suite. Sharing of lessons learned. 2. MEASURING SUCCESS To demonstrate our success, we will openly share our process data comparisons of engineering runs vs lab scale for the 2000L SUB train. 3. BREAKING TRADITION Janssen experience of breaking away from Stainless Steel manufacturing and giving Single Use a well-deserved seat at the table. Finesse CellWorld San Francisco 53

Disposable Cell Culture Process Train Pre Culture 30ml 50L Seed SUB 250L Production -SUB 1000L Harvest -2000L Media -500L Process Vessels 200-500L Finesse CellWorld San Francisco 54

Media Preparation 500L Single Use Mixer (WV >100L) Recirculation Line Powdertainer addition port Liquid addition Lines Media Transfer Train 0.45/0.22 10 Durapore filter 2 x 0.1um 4 Opticap Filters (sterile boundary) 200L Media Storage Bags Finesse CellWorld San Francisco 55

Prove out of probes Electrochemical Advantages Proven Technology Process Range Disadvantages Autoclaved Polarisation Time -DO(~6Hrs) Contamination Risk Optical Advantages Ease of Use Integral to the Bag Good correlation with e chem Disadvantages Technology in early development Limited GMP instances Finesse CellWorld San Francisco 56

DO Probe Trends Batch 1 90 80 70 60 % Dissolved Oxygen 50 40 30 20 10 0 0 2 4 6 8 10 12 14 16 18 Culture Days Batch 1 Reusable DO Batch 1 Single Use DO Finesse CellWorld San Francisco 57

DO Probe Trends Batch 2 % Dissolved Oxygen 100 90 80 70 60 50 40 30 20 10 0 0 2 4 6 8 10 12 14 16 18 Culture Days Batch 2 Reusable DO Batch 2 Single Use DO Finesse CellWorld San Francisco 58

DO Probe Trends Batch 3 % Dissolved Oxygen 90 80 70 60 50 40 30 20 10 0 0 2 4 6 8 10 12 14 16 18 Culture Days Batch 3 Reusable DO Batch 3 Single Use DO Finesse CellWorld San Francisco 59

ph Values 7.5 7.4 7.3 7.2 7.1 7 6.9 6.8 6.7 6.6 6.5 ph Probe Trends Batch 1 0 2 4 6 8 10 12 14 16 18 Culture Days Batch 1 Reusable ph Batch 1 Single Use ph Finesse CellWorld San Francisco 60

ph Values 7.5 7.4 7.3 7.2 7.1 7 6.9 6.8 6.7 6.6 6.5 Single Use ph Evaluation 0 2 4 6 8 10 12 14 16 18 Culture Days Batch 2 Reusable ph Batch 2 Single Use ph Finesse CellWorld San Francisco 61

Cell Culture Suite 2 - Project Milestone Map + Engineering Design + Procurement + Construction + Commissioning + Qualification + Engineering Run Phase 1 Clinical Campaign + Clinical Campaign 14Dec11 Finesse CellWorld San Francisco 62

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