The Use of Disposable and Alternative Purification Technologies for Biopharmaceuticals

Transcription

1 The Use of Disposable and Alternative Purification Technologies for Biopharmaceuticals Tom Ransohoff BioProcess Technology Consultants July 22, 2005 IIR Biopharmaceutical Production Conference

2 Presentation outline Driving forces for use of disposable and alternative purification technologies Examples of current technologies Obstacles to implementation Future possibilities

3 Driving Forces

4 Driving forces for disposable and alternative purification technologies Business drivers for alternative approaches to biopharmaceutical manufacturing: Need to reduce capital investment in high-risk projects Speed to clinical proof-of-concept and commercial launch Operational trends favoring disposables: Increasing focus on cleaning and cleaning validation Increasing availability of suitable disposable processing equipment throughout the biopharmaceutical flowpath: Flexible bioreactors Media and buffer bags Disposable aseptic filling equipment Trend towards large volume monoclonal antibody processes: Increasing productivity and scale of upstream processes Purification increasingly becoming a bottleneck

5 The majority of manufacturing costs for most biopharmaceuticals are fixed costs 100 kg/year 1000 kg/year Overhead/Indirect Labor 22% Utilities & Waste 9% Capital 40% Quality & Indirect 20% Utilities & Waste 6% Capital 29% Materials & Consumables 13% Direct Labor 16% Materials 29% Direct Labor 16% Data from a process economic model of a typical mammalian cell culture-derived monoclonal antibody

6 Project risk and capital cost decrease significantly as development progresses 100% BLA Approval 90% 80% 70% 60% Phase III 50% 40% 30% 20% 10% Preclin Phase I Phase II 0% Time (yr) Probability of Success Cost of Capital * * - Cost of Capital estimate based on discount factors from a risk-adjusted NPV analysis

7 Technologies that reduce capital investment and/or increase speed to risk-reduction milestones are inherently more valuable in high-risk projects There is a compelling financial and risk-management argument for disposable manufacturing technologies in early-stage biopharmaceutical development

8 Examples of Process Use of Disposable Bioreactors Use of disposable bioreactors for production at 100L+ scale: Pierce, LN and Shabram PW, Scalability of a Disposable Bioreactor from 25 L 500 L Run in Perfusion Mode with a CHO-Based Cell Line: A Tech Review, BioProcessing Journal, July/Aug Zhang, S et al, Large Scale Production of Clinical Grade Adenovirus Vectors Introgen s Experience, presentation at WilBio Viral Vectors and Vaccines Conference (2004). Use of disposable bioreactors for production at smaller scales: Jenderek, S et al, Development of a Production and Purification Method for Type 5 Adenovirus, BioProcessing Journal, May/June Use of disposable bioreactors to replace conventional bioreactors in the inoculum train of large scale processes: Knevelman, C et al, Characterisation and Operation of a Disposable Bioreactor as a Replacement for Conventional Steam in Place Inoculum Bioreactors for Mammalian Cell Culture Processes, ACS poster presentation (2002).

9 Disposable products* exist across most of the biopharmaceutical manufacturing flowpath Wave Bioreactors Limited Fermentation Purification Formulation/ Fill Media Prep/ Storage Buffer Prep/ Storage Stedim Celsius Stedim Bags, Hynetics Mixers * - Suppliers listed are examples, not an endorsement of any specific company or technology

10 Downstream processing unit operations Normal Flow Filtration Depth filtration for clarification Nanofiltration for virus removal Sterile filtration Tangential Flow Filtration Ultrafiltration for concentration and buffer exchange Microfiltration for clarification Centrifugation Clarification Inclusion body isolation Cell Breakage/Homogenization For recovery of products expressed intracellularly Refolding For some E. coli products Crystallization/Precipitation Chromatography and adsorptive separations Typical downstream process includes 3 4 chromatography and/or membrane adsorber steps Ion exchange Hydrophobic interaction Affinity Size exclusion Reverse phase

11 Commercial product requirement trends - biopharmaceuticals 2004 Sales by Mfg Technology/Prod Type Mammalian Products Microbial rprotein, $15.4 Mammalian Mab, $ rproteins MAbs Mammalian rprotein, $ Amount Required (kg/yr) N Ave. Sales Price ($/mg) Fastest growing commercial biopharmaceutical segment is monoclonal antibodies at 40%+/year Trend for commercial products is from agonists to antagonists -> larger volume, less expensive ($/g) products Source: BPTC 2004 Annual Manufacturing Capacity Analysis

12 Clinical biopharmaceutical pipeline is weighted towards antibody-based products Pipeline Number of Products Pipeline Fraction MAb-based % Number of Products Mammalian Microbial Percent MAb-based 90% 80% 70% 60% 50% 40% 30% 20% 10% Mammalian Microbial 0 Market BLA Phase III Phase II Phase I Stage 0% Market BLA Phase III Phase II Phase I Stage Biopharmaceutical pipeline is 70% mammalian cell culture Significant trend towards antibody-based products is ~ 85% of mammalian cell culture pipeline This analysis covers recombinant protein and monoclonal antibody product candidates only Source: BPTC 2004 Annual Manufacturing Capacity Analysis

13 Typical material requirements clinical development Pre-clinical development kg Phase I Phase II Phase III kg kg 1 20 kg

14 Material requirements commercial biopharmaceuticals Highly product dependent For microbial fermentation-derived products, commercial requirements range from: ~10 g/yr (e.g., Infergen) to ~5,000 kg/yr (Novolin) Most products require kg/yr For mammalian cell culture-derived products, commercial requirements range from: ~10 g/yr (e.g., Bexxar/Zevalin) to ~750 kg/yr (Rituxan) Most non-antibody products require <10 kg/yr Most antibody products require kg/yr

15 2004 bulk product requirements (kg/yr) mammalian commercial products Total estimated bulk product requirements for mammalian cell culture commercial products (2004): All mammalian products 2,875 kg/yr Monoclonal antibodies 2,810 kg/yr rproteins 65 kg/yr Significant growth in bulk requirements over recent years due to successful MAbs: Bulk Reqmts 2004 (kg/yr) Rituxan Remicade Enbrel Herceptin Avastin Erbitux All others (38) Product Source: BPTC 2004 Annual Manufacturing Capacity Analysis

16 Current Approaches

17 Examples of current disposable format purification products and technologies Membrane adsorbers Pre-packed chromatography columns or cartridges Disposable flow-path system concepts

18 Membrane Adsorbers Remember when... Things change - now Biotechnology Division Sartobind Membrane Adsorbers Matrix: stabilized and cross-linked cellulose >3µm pore size Q,S,C,D and affinity ligands (ProtA, IDA, paba, etc) Very low unspecific adsorption High chemical resistance against solvents,acids and caustic solutions and autoclaving 18

19 Membrane Adsorbers Remember when... Things change - now Biotechnology Division DNA removal clears the DNA below detection limit. J. K. Walter, Boehringer-Ingelheim Pharma in: Bioseparation and Bioprocessing, G. Subramanian (ed.) Wiley VCH, 1998, Vol. II p DNA (pg/mg protein) After Affinity Step After DNA removal step Average of eight 2,000 liter batches using 70 ml Sartobind Average of three 12,500 liter batches using 500 ml Sartobind 19

20 Other examples of process use of membrane adsorbers Martin J, Case Study Orthogonal Membrane Technologies for Viral and DNA Clearance, presented at SCI Membrane Chromatography Conference (2004). Haber C et al, Membrane chromatography of DNA: conformation-induced capacity and selectivity, Biotechnol Bioeng, 88(1) (2004). Slepushkin V et al, Large-scale Purification of a Lentiviral Vector by Size Exclusion Chromatography or Mustang Q Ion Exchange Capsule, Bioprocessing Journal, Sep/Oct 2003.

21 BIOFLASH 12 and BIOFLASH 80 Prepacked Columns Column Specifications Pressure Rating bar w/ module 33 bar Diameters 1.2, 8, and 20 cm Bed Length 5 30 cm Bed Volume 5 ml 5+ L Courtesy of BioSepTec, Inc.

22 High performance packing in a disposable format CM Toyopearl 650 BioFlash 80 (10 cm H) Ret. Time 4.77 Efficiency 823 Assym Efficiency measured using 5% acetone in 0.1M NaCl as mobile phase minutes Courtesy of BioSepTec, Inc.

23 Scale-up of recombinant protein separation on BioFlash pre-packed column 1M NaCl Peak of Interest 300 A MM NaCl 1 cm (D) x 10 cm (H) column 100 1M NaCl A Peak of Interest MM NaCl 100 BioFlash 80 (8 cm D) x 10 cm (H) Purification of EPI-HNE-4 on Macroprep 25S at 100 cm/hr. Ransohoff, T et al, Evaluation of BIOFLASH Prepacked Chromatography Columns for the Large-Scale Purification of EPI-HNE-4 Protein over MacroPrep High S Media, Purdue Chrom. Workshop (1999)

24 Introducing The Millipore Pod Filter Platform Improves process flexibility Decreases processing time More robust and scalable Easier to setup and use Minimizes product loss Enhances operator safety Reduces cleaning requirements Improves process economics

25 Improved Handling and Ease of Use Improved CIP of Hardware Self-contained, disposable Pods Disposable feed ports and fittings No product contact with endplates or process skid Improved Handling No messy spent filters Lightweight, easy to set up and use No hoist or high ceiling required

26 Impact of increasing cell culture titer and scale on downstream processes Cell culture titers of 4 g/l are now achievable for monoclonal antibodies: Wurm F, Production of recombinant protein therapeutics in cultivated mammalian cells, Nature Biotechnology, 22(11) (2004). Smith M, Strategies for the purification of high titre, high volume mammalian cell culture batches, presented at BioProcess International European Conference and Exhibition (2005). New facilities are increasing bioreactor scale to 20+ m 3 to meet requirements of large volume products: Lonza Portsmouth: 3 x 20,000 L (on-line) + 1 x 20,000 L (2006) Genentech Vacaville CCP-2: 8 x 25,000 L (2009) At 4 g/l, a 25,000 L bioreactor will yield 100 kg per batch Increasing production scale and titers present challenges and opportunities for downstream process operation

27 Current approaches to managing high volume processes At 4 g/l, a 25,000 L bioreactor yields 100 kg per batch At a loading capacity of 20 g/l during capture chromatography, 8 cycles on a 2 m (ID) x 20 cm (H) column (CV ~ 630 L) are required At 15 CV buffer per cycle, 75,000 L of buffer are needed per batch per chromatography step Current approaches based on maximizing value of existing conventional technology approaches (cf Smith*): On-line dilution to reduce buffer storage requirements Move to more rigid capture media to achieve higher velocities on chromatography steps Multiple chromatography cycles (5-20) per batch Load chromatography columns at/near break-through Optimize UF steps to improve utilization of membranes These approaches work, but may have a limited lifespan -> parallel track investment in novel technologies * - Smith, M, Strategies for the purification of high titre, high volume mammalian cell culture processes, presented at BioProcess International European Conference and Exhibition (2005)

28 Obstacles and Future Possibilities

29 Potential obstacles to implementation of disposable or alternative technologies Cost Extractables and Leachates Inertia

30 Cost impact of disposable technologies The use of disposable-format purification technology will generally: Reduce capital costs Reduce labor requirements for setup/cleaning/cleaning validation Increase direct materials costs The magnitude of the direct material cost impact for disposable chromatography is assessed for two situations: Clinical manufacturing for an outsourced early-stage product Commercial large-scale manufacturing

31 Typical project costs development and clinical manufacturing (CMO) Clinical Process and Analytical Development Activity Time Approximate Cost Cell Line Development (includes Vector Construction, Transfection, Selection, Identification of Final Clone)* 4 6 months $150, ,000 Master and Working Cell Bank Generation* 4 6 months $75, ,000 Process Development 8 months $500,000 1,500,000 Process Scale-up 3 months $300,000 Analytical Development and Qualification 9 months $150,000 Viral Clearance Validation* (1-3 model virus) 6 12 months $150,000 - $250,000 TOTAL - ~ $1.5-3 million cgmp Manufacturing Clinical engineering and cgmp mfg lots ~$400k 600k per batch (bulk) cgmp aseptic filling* of clinical lots ~$75k 125k per lot * - Activities that are frequently sub-contracted to other service providers

32 Cost impact for disposable chromatography in clinical manufacturing Total Project Costs - Clinical Manufacturing Min Max Process/Analytical Development Costs $1,500 $3,000 Total Manufacturing Costs $475 $2,525 Cost per lot $400 $600 # of bulk lots $1 $4 Cost of DP mfg $75 $125 TOTAL COSTS $1,975 $5,525 Process Assumptions Scale 500 L Titer 1 g/l Yield 50% Output 0.5 g/l Clinical Requirements (g) # of batches 1 4 Media cost of using disposable capture affinity chromatography Assumptions Cost of media $10,000 per L # of cycles per lot 10 Binding capacity 20 g/l Volume of media 2.5 L Cost of use as a disposable $25,000 $100,000 As a % of Total Costs 1.3% 1.8% As a % of Manufacturing Costs 5.3% 4.0% Media cost of using disposable ion exchange chromatography Assumptions Cost of media $1,000 per L # of cycles per lot 10 Binding capacity 30 g/l Volume of media 1.25 L Cost of use as a disposable $1,250 $5,000 As a % of Total Costs 0.1% 0.1% As a % of Manufacturing Costs 0.3% 0.2% With non-disposable approach, initial investment in media will be at least equivalent to single lot cost. Benefits of using disposable chromatography approach may be significant enough in this application to justify modest additional operating costs: Improved manufacturing efficiency Improved process portability Reduced risk of cross-contamination

33 Summary of COG estimates for a typical commercial-scale monoclonal antibody process Mam Cell (Mab) Process Cost v. Scale $ $ $ Cost of Goods ($/g) $ $ $ $ $ $50.00 $ Scale (kg/yr)

34 Cost impact for disposable chromatography in large-scale manufacturing Commercial Manufacturing Costs Min Max Total Manufacturing Costs ($M/yr) $40 $150 Cost per gram $400 $150 Annual requirements (kg/yr) 100 1,000 Total Materials Costs ($M/yr) $5 $44 Materials Costs as % of Total Costs 13% 29% Process Assumptions Scale L Titer 1.5 g/l Yield 60% Output 0.9 g/l # of batches/yr 8 75 Media cost of using disposable capture affinity chromatography Assumptions Cost of media $10,000 per L # of cycles per lot 10 Binding capacity 20 g/l Volume of media per batch L Cost of use as a disposable ($M/yr) $9.0 $84.4 Cost per g product produced ($/g) $90 $84 As a % of Total Mfg Costs 22.5% 56.3% As a % of Total Matl Costs 173.1% 194.0% Media cost of using disposable ion exchange chromatography Assumptions Cost of media $1,000 per L # of cycles per lot 10 Binding capacity 30 g/l Volume of media per batch L Cost of use as a disposable ($M/yr) $0.5 $4.2 Cost per g product produced ($/g) $5 $4 As a % of Total Mfg Costs 1.1% 2.8% As a % of Total Matls Costs 8.7% 9.7% Operating cost impact of disposable chromatography approach far greater in this application. Alternative approaches required to significantly impact costs in large-scale commercial manufacturing

35 Extractables and leachates Extractables and leachates are a potential concern with any disposable technology. Level of concern increases as product purity increases. Weidner presented work at Biogen Idec to address regulatory requests related to extractables: Risk-based assessment of extractables based on Process step Contact time Temperature Solvent Stage of development Vendor provided information on extractables and toxicological testing Conduct extractable tests where warranted based on potential risk Mass transfer principles guide test design Analytical methods for quantification and identification appropriate to situation Results assessed against acceptance criteria or by evaluation of toxicological risk Source: Weidner J Validation Considerations for Disposable Components and Systems, presented at the NC Biotech Symposium, RTP, NC Nov 18, 2004

36 Inertia: driving forces against use of novel technology and approaches There are significant costs and risks associated with process innovation in any highly regulated industry Conservative approach to implementation of new technologies Security of supply is a concern that must be addressed The complexity of biopharmaceutical processes provide additional challenges There is no good time to innovate: significant obstacles to implementation of new technologies exist at every stage of development Result: New technologies often take longer than anticipated to implement, even when a compelling need exisits

37 Future trends Increased use of disposable purification technologies, particularly in smaller volume processes, driven by: Continued advances in and increased use of disposable systems across the biopharmaceutical manufacturing flowpath Implementation of new materials that significantly reduce per liter costs and allow increased throughput, potentially including: Cost-effective affinity media Novel membrane materials and adsorptive separation supports Increasing interest in alternative technologies and operating approaches for large-volume processes as optimization of conventional technologies matures, including: Operationg approaches that move away from big batch and towards continuous purification processes (e.g., SMB-like processes) Integration of unit operations (i.e., semi-continuous capture and clarification) Implementation of novel (to biotech) technologies and unit operations

38 Summary Current existing technology meets some of the requirements for cost-effective disposable purification solutions Membrane adsorbers Pre-packed columns Disposable-format systems Trends in biopharmaceutical manufacturing are increasing the need for disposable or alternative purification technologies: Increasing use of disposable technologies to reduce capital investment and product development times Increasing titer and scale of monoclonal antibody cell culture production The use of disposable and alternative purification technologies will increase as: The trends towards disposable equipment and large volume processes continue New technology and viable solutions are introduced that provide solutions more broadly for purification unit operations

39 Thank you! BioProcess Technology Consultants, Inc. 289 Great Road, Suite 303 Acton, MA

40 Carousel-type SMB evaluation of Protein A separation Columns are mounted on a slowly rotating carousel The columns on the carousel are connected to the pumps and vessels via a rotary valve Switching mode simulates continuous adsorbent flow Courtesy of J. Thommes (Biogen Idec), from Protein A Affinity Simulated Bed Chromatography for Continuous Monoclonal Antibody Purification, Presented at Recovery of Biological Products XI Conference, Banff Canada (2003)