Challenges in the cgmp Manufacturing of hescs: Lessons Learned from Monoclonal Antibodies ISCT 2011 Annual Meeting Rotterdam, The Netherlands May 18 21, 2011 BioProcess Technology Consultants www.bptc.com
Important Considerations in cgmp Manufacturing Capacity Flexibility Quality/ Regulatory Performance/ Productivity Scalability Speed Cost
Annual Sales ($ Billions) Growth in Monoclonal Antibody Product Sales 45 40 35 30 Largest class of biologic products today 25 20 15 10 5 0 2002 2003 2004 2005 2006 2007 2008 2009
Hurdles in Monoclonal Antibody Development Immunogenicity HAMA limited early product development Productivity Low titers and poor purification yields caused initial high COGS Product Consistency Reproducible glycosylation with scale and process
Evolution of Monoclonal Antibody Products First murine MAb approval Centoxin approved but withdrawn from market First IgG-fusion protein approval 1976 1986 1991 1997 2000 2002 Köhler & Milstein discover MAbs First Chimeric MAb approval Zenapax First humanized MAb approval First fully human MAb approval
No. Products Approved Approval of Monoclonal Antibody Products by Year 6 5 4 3 2 1 0
Standardized Production Host and Culture Conditions SP2/0 NS0 CHO Chemically defined, serum (or protein)- free media Operating parameters improve productivity to 50 pg/cell/day CHO adapted for suspension culture Fed-batch culture with specific nutrient feeds Increased peak cell densities and run duration
Titer (mg/l) Typical Monoclonal Antibody Cell Culture Process 10 100 ml vial 50 L 200 L 1,000 L 7,500 L 20,000 L High cell density and high growth rate media enable maximal seed density in inoculum train and production bioreactor Cell densities at harvest 3 x 10 10 cells/l Titers 5 g/l in 12 15 day culture Culture time from vial to harvest ~22 25 days 11000 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 0 2 4 6 8 10 12 14 16 18 20 22 Days
Titer in mg/l Innovation has Increased Titers and Yields 10,000 1,000 100 10 1 New technologies to improve cell line development and expression levels coupled with improved and optimized media, supplements, and bioreactor conditions have increased titers Over 3 log increase in titer over the last 20 years Current commercial products range from 0.2 3.0 g/l 1980 1985 1990 1995 2000 2005 2010 2015 2020 Antibodies Recombinant Antibodies Hybridoma Fc Fusion Ref: W. Noe (2011), T. Charlebois (2006), M. Smith, (2005), F. Wurm (2004)
Platform Processes Enable Large Scale Purification Ref: B. Kelley, ACS Conference, (2006)
Cost of Manufacturing Monoclonal Antibodies Phase of Development Quantity Required (Kg) Relative Production Scale Estimated Cost 1 0.5 2 1 2 1 10 1 3 3 5 25 3 5 $0.75 1.2 MM Per batch $1.8 2.5 MM Per batch $2.5 5.0 MM Per batch Commercial 10 1,000 (per year) 5 10 $1,000/gram Typically $150 Early stage production typically priced on per batch basis due to small scale and incomplete process development Batch prices similar at each scale but larger batch size lowers COGS Commercial production at 2,000 20,000 L Higher titers lowering average commercial scale bioreactor requirements
Cost $/gram Large Scale Monoclonal Antibody Production Costs $500 Depreciation $400 $300 19% 9% 13% 7% 52% Raw Materials Consumables Labor Misc. $200 $100 $- 0 3 6 9 12 MAb Titer g/l Based on annual production of 100 Kg/year; bioreactor size ranging from 1,000 10,000 L
Issues in the Manufacture of Stem Cells Product complexity greater than monoclonal antibodies Final product not homogeneous May contain partially differentiated or undifferentiated cells Induced pluripotent stem cells are genetically modified Manufacturing challenges Anchorage dependent growth requires microcarriers Production scale limited due to metabolic concerns, low density propagation, and limited experience Wide variety of culture systems currently in use which may have limited scalability Unique supply chain issues Fragility of cells may require frozen storage and shipping Limited shelf-life of product Shipping protocols must ensure product viability and sterility
Number of Products Allogeneic Cell Therapy Products in Development 60 50 40 30 20 10 0 Market Phase 3 Phase 2 Phase 1 Preclinical Embryonic Somatic Includes company sponsored products only. Several investigator -sponsored trials in progress
As With Early Biologics, Process = Product Product Complexity Will require extensive testing, especially early in development, to ensure product quality and consistency Potential for different advice or requirements by individual regulatory authorities Process Changes Lack of product and process knowledge limit ability to make changes during development Manufacturing process should be fully cgmp compliant and relatively final prior to Phase 1 Product Quality Concept very different than monoclonal antibodies Identity testing requires use of expression markers
Critical Factors in Cell Therapy Manufacturing Quality of all product contact surfaces and media/buffers Product contact surfaces can impact differentiation, changing product composition Different microcarriers have different impact on cell growth and differentiation Control of product to prevent contamination Viral inactivation or removal not possible Terminal sterilization not possible Operator training essential Potency of growth factors used to initiate differentiation Often several factors used to initiate differentiation Balance of activities essential to initiate correct differentiation Limited material available for QC testing
Cell Therapy Manufacturing Process WCB (undifferentiated hecs) Cell Expansion Differentiation Cell Harvest Formulation and Fill Cryopreservation Ref: www.geron.com
Cell Therapy Process Parameters Typical Harvest Densities 2D culture in T-flasks 8.0 x 10 5 cells/cm square 3D culture in bioreactors un-optimized 2.0 x 10 9 cells/l 3D culture in bioreactors optimized 2.0 x 10 10 cells/l 50% overall recovery yield from bioreactor to vial Low dose applications, e.g., spinal cord injury 2.0 x 10 7 cells/dose; single dose High dose applications, e.g., congestive heart failure 5.0 x 10 8 cells/dose; single dose Target patient population of 10,000 550,000 patients at market peak Market penetration of 30 50% at market peak
Very Small Production Volumes Required Low dose/small patient population/ Low market penetration Total annual production requirement 6.6 x 10 10 cells Total annual production volume 6.6 L Production in T-flasks or spinners High dose/high patient population/ High market penetration Total annual production requirement 1.4 x 10 14 cells Total annual production volume 14,000 L Production in small bioreactors 15 batches/yr @ 1,000 L/batch 19
Potential Commercial COGS for hesc Products Case 1 Case 2 Case 3 Bioreactor size (L) 500 1,000 2,000 Cell density at harvest (cells/l) 2.0 x 10 10 2.0 x 10 10 2.0 x 10 10 Overall yield 50% 50% 50% Total cells per batch (cells) 5.0 x 10 12 1.0 x 10 13 2.0 x 10 13 Doses per batch (5 x 10 8 cells/dose) 10,000 20,000 40,000 Batches per year 30 15 8 Doses produced per year 300,000 300,000 320,000 Cost per batch ($ million) 11.74 8.41 11.53 Cost per dose ($/dose) 1,174 420 288 Case 1 & 2 = Single use bioreactor; Case 3 = Stainless steel bioreactor Costs based on hypothetical manufacturing process and do not include storage or shipping of product Cost predominantly driven by depreciation and raw materials
Single Use Bioreactors for hesc Manufacturing Xcellerex XDR TM Bioreactor Sartorius Stedim Biostat Culti-bag Thermo Fisher (Hyclone) Single-use Bioreactor GE Healthcare Wave Bioreactor ATMI Nucleo TM Bioreactor
hesc Manufacturing Facility Supply Corridor Inoculum Class C Areas: Inoculum preparation Wash Materials Corridor Staging Area Media & Buffer Preparation Upstream Processing Fill/finish Class D Areas: Cell culture (Single use bioreactor) Downstream processing Media and buffer preparation Fill/Finish Cell processing Personnel Corridor Cell recovery Clean glassware unloading and storage Media and buffer storage in controlled, non-classified areas Process Area 6,781 ft 2 Total Area 13,014 ft 2 Class C 667 ft 2 Class D 3,315 ft 2 Process Equipment $5.0 MM Total Capital Cost $15 MM
Thank You! Howard L. Levine, Ph.D. hlevine@bptc.com +1.781.281.2703 BioProcess Technology Consultants, Inc. 12 Gill Street, Suite 5450 Woburn, MA 01801
Follow us www.bptc.com www.bioprocessblog.com bptcglobal www.linkedin.com/company/bioprocess-technology-consultants-inc.