Process Development for Cell Therapy and Viral Gene Therapy
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Today’s viral vector manufacturing processes remain challenging. Process development is a critical enabler to bring safe, effective, sustainable products to market to address patient needs. When done properly, it can reduce the timeline of the project and the cost of producing the therapeutic product. The webinar discusses our strategies for developing lentivirus and adeno associated virus (AAV) and the impact these early decisions can have on commercial readiness. Watch the interactive webinar now: https://bit.ly/2VplwQq
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Process Development for Cell Therapy and Viral Gene Therapy
- 1. Merck KGaA Darmstadt, Germany Process Development for Cell Therapy and Viral Gene Therapy Elie Hanania Head of Process Development
- 2. The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada. 2
- 3. Viral Vectors are vehicles used to deliver a genetic payload into a target cell Key Feature Include: Viral Vectors in Cell & Gene Therapy Safety lacks viral genes involved in replication Value targeted delivery for optimal efficacy Manufacturing capable of production and purification to high levels Applicability infect/transduce a wide spectrum of cells Environmental Containment controlled virus dissemination 3
- 4. Generalized Viral Vector Manufacturing Process Thaw and Expand Cells Seed Expansion Vessel Vector Production –Infection/ Transfection Filter Clarification Concentration and Final Formulation Sterile Filtration Fill and Finish • Volume • Media • Virus Size & Properties • Purification Mode • Nature of Contaminants • Pressure / Shear • Final Filtration • Aggregation 2nd Chromatography Step • Volume • Surface Area • Pore Size • Flow Rate • Pressure /Shear • Virus Stability Factors to consider Factors to consider Downstream Midstream • Cell type • Final titer • Adherent/ suspension • Pressure /Shear • Virus Stability Factors to consider Upstream Lysis / Benzonase® Concentration and Diafiltration 1st Chromatography Step 4
- 5. Industrialization ongoing, but multiple solutions exist on market TFF UF/DF Pellicon® 2 Cassette Biomax® 100 kD Membrane DNA Digestion Benzonase® Endonuclease ELISA Kit II Mobius ® Mixer 1° Chromatography Fractogel® TMAE Resin Media & Inoculum Prep Millipore Express® HPF > SHR Filters Cell Growth & Virus Production Mobius® Bioreactor Novaseptum® Sampling Aervent® (vent filter) Virus Harvest (Cell Lysis) Mobius ® Mixer 1° Clarification Polygard ® CR Filter Clarisolve® Filter Millistak ® Filter TFF UF/DF Pellicon® 2 Cassette Biomax® 300 kD Membrane Sterile Filtration Durapor ® 0.22mm Membrane 2° Clarification Clarigard ® Filter Milligard ® Filter Polysep® II Filter Durapore® Membrane 2° Chromatography Fractogel® DMAE Resin F&F Mobius® Integrated Fill Finish Solutions Thaw and Expand Cells and Seeds 5
- 6. Raw Materials and Reagents Equipment Consumable Sets Single Use Technology Path to Commercialization – Critical Factors Viral Vectors Manufacturing Product Quality Product Yield Titer Purity Equipment Unit Operations Mode of Operation Stability Tolerance Feasible Range Process Time 1 cGMP Compatible 4 Robust 2 Scalable 3 Reproducible 6
- 7. Master Cell Line Production (MCB); 800 vials Limited time factoring in release testing No Process Development work done Requirements exceeded typical 200 vials Cell expansion performed in adherent cultures in ten-layered CellSTACKS All components, reagents, supplies procured Batch records approved Scenario 1: Lack of Process Development 1 cGMP Compatible 4 Robust 2 Scalable 3 Reproducible Solution • Go back to Process Development • Optimize cell processing; focus on duration • Assess cryopreservation conditions • Confirm recovery & viability post thaw • PD work added 10% to total cost • Time Impact: None Outcome • Low recovery and viability upon thaw • Financial impact: Additional 35% of total cost • Time Impact: Additional 24% of total duration 7
- 8. “Rushed” Production and Purification Tech transfer to Process Development (PD) Process required development and verification at scale Limited Process Development performed Client rushed into cGMP production. Changes were made during the run - NOT tested in PD Changes included: Cell line, production parameters, size of purifications platforms Scenario 2: Lack of Process Development Solution • Extended PD with scale up verification studies • New cell line tested in PD • Production parameters re- assessed and locked for Tech Transfer • Based on scale run, filters, TFF cartridges & Chromatography columns were scaled appropriately • Engineering run was successful • Additional PD work added 10% to the total cost • Time Impact: Additional 15% of duration to deal with project creep Outcome • Low titer and yield; material not enough for client needs • Financial impact: Additional 53% of total cost • Time Impact: Additional 31% of total duration. Extended time to close events and initiate CAPAs 1 cGMP Compatible 4 Robust 2 Scalable 3 Reproducible 8
- 9. Step-wise Planning for Production and Purification Client started with tech transfer to PD Process performed for verification Process was modified for cGMP production. Progressive scale up studies done Client transferred analytical assays and requested development of others Assays qualified in preparation for cGMP runs Batch records were generated based on tech transfer from PD to GMP Engineering run performed. Production summary review Scenario 3: Process Development Completed Outcome • Time and budget for thorough tech transfer and development activities • Tasks fulfilled as planned • High yield; material exceeded client needs • Financial impact: None. Scope well defined and priced accordingly • Time Impact: None. Project timeline discussed with client. Updates provided during the PD 1 cGMP Compatible 4 Robust 2 Scalable 3 Reproducible 9
- 10. Process optimization critical to maximizing yield Lentivirus production processes remain challenging 67-100% Step yields up… ~55% Benzonase® Endonuclease/ Clarification TFF1 TFF2 Sterile Filtration Chromatography 45-50% ~85% 30-85% Up to 10-20% 10
- 11. Process development levers exist to maximize yield AAV production processes achieving higher process yields 83-100% Step yields up… 60-100% Benzonase® Endonuclease/ Clarification TFF1 TFF2 Sterile Filtration Chromatography 50-90% ~85% 70-93% Up to 20-40% 11
- 12. Levers for optimizing unit operations Step Primary Focus Optimization Impact Tangential flow: Concentration Shear Flow rate Control pressure Functional titer Tangential flow: Diafiltration Virus Stability Buffer composition Number of dia-volumes Yield and functional titer Chromatography: Ion Exchange Recovery and Purity Adjust conductivity Adjust pH Load, wash, elution rates Titer, yield, residuals, impurities Functional titer Chromatography: Affinity Recovery and Purity Adjust pH Elution Buffer composition Titer, yield, impurities Chromatography: Size Exclusion Purity Operation rate Choice of resin Yield, impurities Filtration Step Recovery Virus Stability Pressure and rate Membrane type and size. Titer, yield, aggregation, Functional titer 12
- 13. Early engagement accelerates best practice sharing PD and CMO manufacture critical on path to commercialization Client Key Activities Stage Review | Stage Review | Stage Review | Stage Review | 3 months 3 months for first batch 3 months Tech Transfer: 3 months Optimization: 6-12 months 13
- 14. Characterization is critical to ensure safety, consistency and robustness Missed Opportunity: Characterization Early (PD) Late (Eng / GMP) Characterization studies reduce risk and delays when done in parallel in PD • Mass Balance • Process appropriateness • Material/supplies appropriateness • Scale assessment ProductProcess • Safety • Reproducibility • Trending • Contaminants • Residuals • Potency • Stability • Identity • Appearance • Titer • Infectious titer • Purity 14
- 15. Value of templated process 1 2 3 4 5 6 7 8 9 101112 1 2 3 4 5 6 7 8 9 101112 1 2 3 4 5 6 7 8 9 101112 Clinical Activity Release to clinic Limited Extensive GMP1 Confirmation BOM/BR Testing Gap/Risk Analysis Engineering Tech Transfer Process Development GMP1 Eng. Run Clinical MCB Release to clinic Limited WCB Confirmation Testing PD Cell Bank Templated processes speed path clinical material and improve quality through standardization Largest benefit when leveraging production cells through downstream processing, avoiding: Cell banking activities Tech transfer / most of process development Engineering run Custom Bill of Material / Batch Record creation TemplateCustom Potential savings: 14-18 months15
- 16. • Industrialization of viral vector manufacturing is needed and in progress • For successful commercialization, four factors must be considered during process development: 1. cGMP Compatible 2. Scalability 3. Reproducibility 4. Robustness • Templated processes speed path clinical material and improve quality through standardization Summary Slide 16
- 17. elie.hanania@milliporesigma.com Elie Hanania The vibrant M, Aervent, Benzonase, Biomax, Clarigard, Clairisolve, Durapore, Fractogel, Milligard, Millipore Express, Millistak, Mobius, Novaseptum, Pellicon, Polygard and Polysep are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources. © 2019 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.
Notes de l'éditeur
- Stage gate reviews will include the team as well as the client. These serve as an interim “health check” to assess the status and risk of the project. Additionally, each stage exit includes a management review. Client’s Product Development cycle is included to help align our nomenclature and definitions.