Disposable Technologies for Purification of Biopharmaceuticals

Transcription

1 Disposable Technologies for Purification of Biopharmaceuticals Thomas C. Ransohoff BioProcess Technology Consultants, Inc. NE Society for Industrial Microbiology Winter Meeting Boston, MA December 14, 2009

2 Outline Why Disposables/Single Use? Single Use Technology Overview BioSMB An Enabling Disposable Purification Technology

3 Why Disposables?

4 Definition of Biopharmaceuticals Biologic Products are products that are made by or composed of viable organisms or biopolymer analogs Recombinant Proteins Monoclonal Antibodies Natural Hormones and Enzymes Synthetic Peptides and Oligonucleotides Antibiotics, Plant & Animal Extracts, Allergens Vaccines Gene Therapy Products, Human & Xenogenic Cells & Tissues Blood & Blood Derivatives, including polyclonal antibodies

5 Biopharmaceutical Industry Growth Biopharmaceutical Commerical Product Sales Growth 40 Annual Sales ($B) Mammalian Recombinant Products 109 Kg required for 2008 Mammalian MAb Products 6,918 Kg required for 2008 Microbial Recombinant Products 12,975 Kg required for 2008 Microbial MAb Products 4 Kg required for BPTC database covers 126 commercially marketed biopharmaceuticals as of 2009

6 General Scheme for Biopharmaceutical Bulk Drug Substance Processes Intracellular (microbial fermentation) Working Cell Bank Bioreactor Conversion Extracellular (microbial fermentation and mammalian cell culture) Upstream Process Cell Harvesting Cell Disruption/Refold Cell Removal Isolation/Recovery Isolation/Recovery Downstream Process Purification Purification Bulk Formulation Bulk Formulation

7 20,000 L Fermentation Suite Source: Lonza Presentation, US Operations Overview

8 Purification Large-Scale Chromatography Source: Lonza Presentation, US Operations Overview

9 Biopharmaceutical Development Clinical PD and Mfg Timeframe Preclinical 30-Day Wait Clinical Development BLA Review Product Marketing Product Concept Development Laboratory & Animal Studies Basic Research R&D Reg. Strategy Product & Mfg. Development Finalization Short Term Long Term Treatment IND IND Submission IND Preparation Initial Review Supplemental Reporting & Review Human Studies Phase I Phase II Phase III Phase IV? BLA/EMEA Submission RTF / Non-approval Drafting Review & Approval Supplements Manufacturing Benchtop Pilot Clinical Commercial Secondary Time 1-5 yrs (2.3yrs) 0.5-2yr 1-3yrs 1-5yrs (total avg. 3-6 yrs) 1-3yrs (1.8yrs.) Costs $1-50 Million $ Million $ Million Adapted from: G. Gamerman, Seraphim total 5-12yrs (7 yrs.)

10 Timing of Plant Construction Phase I (12 months) Safety Plant investment decisions must be made long before product approval Phase II (24 months) Dose Finding First Efficacy Clinical Development Timeline (6-7 years) Phase III (24 months) Pivotal Trials Filing & Review (18 months) Lead-Time for Building a Commercial Plant (~4 years) Design (12 months) Construction (24 months) Validation (12 months) Product Launch Source: P. Seymour, IBC Bench to Clinic 2002

11 Mammalian Cell Culture Facility Costs Capital Cost per Liter vs. Plant Capacity $30,000 $25,000 Cost per L (000) $20,000 $15,000 $10,000 $5,000 $ Log Plant capacity (L)

12 Pipeline Weighted Towards MAb Products Monoclonal antibodies represent the fastest growing segment of the pharmaceutical industry 85 90% of the mammalian cell culture product pipeline Approximately 65% of all biopharmaceutical products in development are produced in mammalian cell culture No. of Products Mammalian Microbial Other (Plant, Insect, etc.) Percent MAb-Based 100% 80% 60% 40% 20% Mammalian Microbial 0 Market BLA/NDA Phase 3 Phase 2 Phase 1** 0% Market BLA/NDA Phase 3 Phase 2 Phase 1**

13 Accelerating Speed to Clinic Y1 Y2 Y3 Activity Q1 Q2 Q3 Q4 Q1 Q2 Q3 Q4 Q1 Q2 Q3 Q4 Pre-clinical stage 1 Cell line development Analytical development Process development MCB Prep/Test Reference Standard Development and Characterization Clinical formulation development Pre-IND meeting DS Mfg (Engineering and GMP clinical runs) and release testing DP Mfg and release testing Stability studies (DP and DS) IND filing MILESTONE POINT 1 - IND Filing Platform Processes Disposable Technologies

14 Project Risk and Cost of Capital are Significantly Higher for Clinical Manufacturing 100% 90% 80% BLA Approval 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

15 1) Technologies that reduce capital investment and/or increase speed to risk-reduction milestones are inherently more valuable in high-risk projects 2) CMC is often on the critical path in early-stage biopharmaceutical development There is a compelling financial and risk-management argument for disposable manufacturing technologies in early-stage biopharmaceutical development

16 Driving Forces for Single-Use Technologies Improved return on capital Reduced and deferred capital investment Increased speed of deployment Process control and portability Process and product flexibility Improved ability to manage and implement change

17 The Biopharmaceutical Facility of the Future Facility design will incorporate high titer (>10 g/l) processes Facilities of the future will require greater DSP space and capabilities to better handle the high titer bioreactor output Ratio of bioreactor space to DSP space will decrease Use of disposable technologies can reduce capital investment by over 50% and operating costs of manufacturing facilities (Roebers, 2009) Smaller bioreactors will produce similar quantities to today s larger bioreactors Smaller facility requirements may enable smaller companies to construct and manage their own facilities more cost effectively Roebers J, Future trends in biopharmaceutical operations and facilities, presented at BPI 2009, Raleigh NC

18 Single-Use Technology Overview

19 Increasing Scale of Disposable Bioreactor Offerings Vendor Scale Product Name Technology Xcellerex Up to 2,000L XDR TM Bioreactor Stirred tank Thermo Fisher (Hyclone) Up to 1,000L Single use Bioreactor (S.U.B.) Stirred tank GE Healthcare (Wave) Up to 500L Wave Bioreactor Rocking Platform

20 Disposable Products* Now Exist Across the Entire Biopharmaceutical Manufacturing Flowpath GE (Wave), Thermo Fisher (Hyclone S.U.B.), Xcellerex (XDR) Fermentation Purification Disposable Sensor Technology Formulation/ Fill Media Prep/ Storage Buffer Prep/ Storage Sartorius Stedim (Celsius), Flexicon (DAFPA) Sartorius Stedim, Thermo Fisher, ATMI, TNTC * - Suppliers profiled are not a complete list nor an endorsement of any specific company or technology

21 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

22 Disposable Format Depth Filtration: Improvements in Hardware Design 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 Courtesy of Millipore

23 Disposable format purification product examples: Chromatography and Adsorptive Separations Membrane adsorbers Sartorius Sartobind Pall Mustang Disposable chromatography columns GE Ready to Process TM BioFlash TM Other Scouting columns and technologies * - Suppliers profiled are not a complete list nor an endorsement of any specific company or technology

24 Membrane Adsorber Use in Commercial DSP...clears the DNA below detection limit 200 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 Courtesy of Sartorius From Clone Commercial

25 Other Literature 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 Vogel J et al, Industrial Scale Membrane Chromatography for Rapid Capture of Complex Protein Drugs from Continuous Perfusion Culture, presented at Recovery of Biological Products 13 (2008).

26 Disposable Chromatography Technology Examples: Pilot/Process Scale Ready-to-Process Chromatography Columns (GE Healthcare) 12.6, 25.1, and 35.9 cm ID x 20 cm H BioFlash Disposable Format Chromatography (DFC TM ) Columns (BioFlash Parnters) 1.2, 8, and 20 cm ID x variable H

27 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 Example of emerging TFF technology: Sius (Novasep); Reverse SPF (Pall); phase Petrone J, RXIII Conference Example of emerging Centrifuge technology: Centritech (Pneumatic Scale)

28 BioSMB: An Enabling Disposable Purification Technology

29 The Biopharmaceutical Facility of the Future Plant has 6 x 2,000 L bioreactors (possibly single use bioreactors) 12 day fed batch CHO culture for MAb Production 2,000 L volume, 10 g/l = 20 Kg MAb in harvest 80% purification yield = 16 Kg per batch Harvest every 4 days 85 harvests/year (340 days) = 1,360 Kg/year Capital investment < $100M Overall COGS < $70 per gram

30 Disposable Format Availability

31 Status & History of SMB Technology SMB and Continuous Chromatography are well established unit operations Advantages: Reduction in buffers & solvents (40 60%) Reduction in resin volume (40 60%) History of SMB: SMB developed for petrochemical industry First applications in food industries Fine chemical industries applications Chiral separations for pharma industry Biopharmaceutical applications

32 Multicolumn Countercurrent Concept v Wash Elution Wash

33 SMB Technology: Market Place Static Type SMB UOP NovaSep Mitsubishi Puritech Carousel Type SMB Calgon Carbon Corporation SepTor Technologies

34 BioSMB TM Operation Wash Elute Clean Equil. Feed Prod. Waste Disposable valve cassette

35 BioSMB TM Operation Wash Elute Clean Equil. Feed Prod. Waste Disposable valve cassette

36 BioSMB TM Operation Wash Elute Clean Equil. Feed Prod. Waste Disposable valve cassette

37 Tarpon s BioSMB Technology Platform Continuous Disposable Multicolumn Chromatography SMB made for Biopharm applications Fully disposable fluid path including valving Flexible configuration modular design Multi column or Multi device mode with unlimited scale Fully disposable-format separation solution Scalable from bench to large-scale manufacturing

38 BioFlash DFC TM Cartridges Features: Various sizes available Biocompatible & sanitary Fully prepacked & tested Wide range of bioprocess separation media possible Consistent high performance Designed for cleanability to allow multiple cycles

39 BioSMB Applications Fully disposable processing train Paired with disposable bioreactors (up to 2,000 L) Clinical manufacturing of MAbs Full scale production for some rprotein therapeutics and MAbs Vaccine manufacturing Large scale processes (5,000 20,000 L bioreactors) Substantial reduction of purification and total manufacturing costs Smaller footprint process and support systems such as buffer tanks, WFI (More product purified per sq ft manufacturing space) Facility fit: enable newer high titer processes to fit inexisting facilites Smaller, less expensive, more flexible downstream processing Increased titer: add more disposables to the same hardware

40 Facility of the Future: Disposable Protein A Step Clinical Manufacturing of MAb (Protein A step) * Bioreactor volume: 2000 L Expression level: 5 gm/l Protein A media: MabSelect SURE ** Pre packed in BioFlash format Batch BioSMB Column Volume No. of columns Cycles per Batch Column diameter 71 L cm 2.5 L cm Column height 25 cm 6cm * Assuming 48 hr processing time for the Protein A chromatography step ** GE Healthcare Product packed into 8 cm Diameter X 6 cm Height BioFlash format Bioflash DFC cartridges (8 cm and 20 cm ID shown) * Bisschops, M. Disposable Format Downstream Process using Continuous Multi-Column Chromatography, IBC Single-Use Conference, Jun 2009.

41 BioSMB Opportunities Proof of principle has been delivered using: Prepacked columns (Bioflash DFC ); Monolithic cartridges (BIA Separations CIM); Expanded Bed Adsorption (Upfront Rhobust ); Membrane Adsorbers (Undisclosed). Protein A chromatography; IX chromatography; HIC chromatography; Mixed mode chromatography; Gel filtration chromatography.

42 Conclusions Product and process innovations resulting in higher yields per batch and lower demand for bioreactor capacity implies: Investments in manufacturing facilities will continue to slow Disposable/single use technologies possible for some commercial supply Single use, disposable format purification technologies are increasingly available to support output from disposable bioreactors All conventional DSP unit operations now have disposable format solutions Technologies such as BioSMB offer potential to address commercial disposable purification requirements

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