BPI.Seminar Semina opelmg ropelmg.com com

Transcription

1

2 Send questions to:

3 Moving to the next level of Manufacturing Oct 14 th 2009 BPI Raleigh Rolf Douwenga VP Global Research and Development DSM Biologics. DSM CONFIDENTIAL

4 FL15 Agenda Introduction ti Industry Issues How does DSM meet these challenges XD Process technology DSP 4

5 DSM Pharmaceutical Products Global Presence Biologics: Groningen, NL (mammalian) Capua, It (microbial) Corporate Office Parsippany, NJ USA Research & Development: Delft, Netherlands Cambridge, MA, USA Geleen, Netherlands 5 Chemical: Linz, Austria Venlo, Netherlands Regensburg, Germany Secondary: Greenville, NC USA

6 The Future of Protein Manufacturing TODAY -Large Stainless Steel Bioreactors -DSP Bottlenecks -Large Footprint / High CAPEX TOMORROW -Disposable Systems -Efficient Processes -Small Footprint / Low CAPEX Photograph by courtesy of Lonza 20,000L Stainless Single Use Bioreactor w/xd Technology Steel Fed-Batch Bioreactor 6

7 DSM Biologics Mission A premier provider of manufacturing technology & services to the global biopharmaceutical industry Manufacturing Services Technology Solutions CHO XD : Extreme Density PER.C6 PER.C6 :HumanCellline Cell-line NS0 DSP : Technologies. 7

8 DSM Biologics Mission A premier provider of manufacturing technology & services to the global biopharmaceutical industry Manufacturing Services Technology Solutions CHO XD : Extreme Density PER.C6 PER.C6 :HumanCellline Cell-line NS0 DSP : Technologies. 8

9 What is XD Process Technology? Extreme Density Cell-culture: DSM proprietary process technology Boosts titer/bioreactor productivity 5-10 fold Boosts cell density 5 10 fold Maintains very high cell viability >90% Good Product Quality Applicable to many cell-lines (incl. CHO and PER.C6 ) Batch time days Single concentrated batch of product Also for existing processes!! 9

10 XD Features Cell Culture Mode of Manufacturing Fed Batch Feed concentrate Build up Metabolites Osmo increase Changing environment Reducing cell viabilities Concentrated Harvest Batch identification Perfusion Medium Feed Wash out Metabolites No Osmo increase Constant environment High cell viabilities Dilute harvest Large harvest 10

11 XD Features XD Process Cell Culture Mode of Manufacturing Fed Batch XD Perfusion Feed concentrate Build up Metabolites Osmo increase Changing environment Reducing cell viabilities Concentrated Harvest Batch identification Medium Feed Medium Feed Wash out Metabolites Wash out Metabolites No Osmo increase No Osmo increase Constant environment Constant environment High cell viabilities High cell viabilities Concentrated Harvest batch identification Dilute harvest Large harvest.combining the best of both worlds 11

12 XD The Process Fresh Medium Spent Medium Cells and Product Refine Technology, LLC Alternating Tangential Flow (ATF) System Image from a e ts 12

13 XD - High Cell Density Unmatched Biomass Cell density (x10 6 cells/ml) XD Fed-batch Dry cell weight (g/l)

14 XD - PER.C6 Results g/L in total broth (40g/l in supernatant) t) Titer (g/l) Yoeld g/l mln cells/ml 4L XD Pro I 4L XD Pro II 4L XD - Pro II 4L XD - Pro III Time (d) PER.C6 cell line is used as example 14

15 50 L XD scale-up runs mln 100 Cellprofile 2 L & 50L XD runs Consistent high titers and cell density at 50 L scale: 0E6 cells/ml) VCD ( Time (day) Titer: 20 gram/l at day 12 VCD: 180 mln cells/ml g/l IgG (g/l) VCD 2 L PCM VCD 50 L PCM Titer profile 2 L & 50L XD runs Equipment works well!! Optimized XD SUB Automatic ATF Improved aeration Time (day) IgG 2 L PCM IgG 50 L PCM 15 2L and 50L runs comparable

16 XD - CHO case-1: Boost of cell density and viability High viability >90% 90 VCD (x 10e6 cells/ /ml) VCD-XD Viab-XD VCD-Fed batch Viab-Fed batch Boost of viable cell density >120 mln/ml Viability (%) Time (days) Scale of the XD process: 3L bioreactor (2 L working volume) Medium used: CD-CHO medium (without any optimization/fortification for XD ) Cells grow up to x10 6 cells/ml Viability always stays high ( 90%) 16 DSM CONFIDENTIAL

17 XD -CHO case 1: Titer and productivity i boost of existing i fed-batch process 10 9 Maximum titer = 8.8 g/l 8 XD Fed batch Volumetric productivity = 0.27 g/l/d Specific productivity = ~14 pcd Pro oduct titer (g/l L) Boost of titer: From 1.2 to 8.8 g/l 2 1 Maximum titer = 1.2 g/l Volumetric productivity = 0.06 g/l/d Specific productivity = ~14 pcd Time (days) Compared to the fed-batch, XD delivers 7.3-fold more product Titer and volumetric (bioreactor) productivity get a boost 17 DSM CONFIDENTIAL

18 XD -CHO case-2: Titer and productivity boost of existing Perfusion process Maximum titer of around 13 g/l - Specific productivity: pcd - Volumetric productivity: ~0.9 g/ld Viability around 70% 10 XD IgG con ncentration (g/l L) Spin-filter perfusion Viability starts below 90% Boost of Titer and productivity 2 Trigger to start Production - Maximum titer of 0.4 g/l - Specific productivity: pcd - Volumetric productivity: ~0.2 g/ld Typical break up of a fed-batch process Time (days) Compared to the perfusion, XD delivers much more concentrated product 18 Titer and volumetric (bioreactor) productivity get a boost

19 XD -CHO case-2: High viability until the end of the batch 70 High viability (90-98%) XD-Increased flow (10-20%) VCD (10 6 cells/m ml) XD-Standard flow Range of normal FB cell densities Typical break up of a fed-batch process Time (days) Cell line: CHO Scale of the process: 2L bioreactor (1 L working volume) Medium used: customer-specific medium (without any optimization/fortification for XD ) Cells grow up to *10^6 per ml 19 Viability stays high for most of the run (apoptosis occurs at day 16-17)

20 XD Media Consumption with PER.C6 PER.C6 C6 XD TM Medium Consumption Profiles dium Con nsumption n (v/v) Me Optimization Clone 35 6 months 30 Optimization Media consumption 25 1 year PER.C6, IgG, PCD Product Titer (g/l) At harvest After Days 1 L XD "Classic Process" 4 L XD - New Clone 4 L XD - New Clone, Improved Mk I Process 4 L XD - New Clone, Mk II Process 4L XD - New Clone, Mk II.2 Process 4L XD - New Clone, Mk II.2 Process Run 2 4L XD - New Clone, Mk II.3 Process 20

21 XD - Product Quality XD samples are comparable to reference fed batch material in terms of purity, charge heterogeneity, aggregation level, bioactivity and glycosylation profile. N -Glycan profiling 60 % Fed batch XD 1 mid XD 1 late XD 2 mid XD 2 late GO G1 G2 glycoform 21

22 XD - A new Life Cycle concept Clinical Development Launch Maturity 50 L XD 250 L XD Multiple 250 L XD KG/batch CT I/II supply KG/batch CT III/comm. supply KG/batch Comm. supply >20 Kg products Flexible integrated processes, using disposables; Larger production volumes already at smaller scales, low CAPEX, small foot print Copy the Process Concept after launch (quick adjustment of capacity to your needs) 22

23 Example DSP technology to purify XD harvests. EBA Expanded Bed Adsorption A Close cooperation between DSM and Upfront Chromatography A/S 23

24 EBA - Expanded Bed Adsorption ρ=15 kg/l! Direct Capture (no filter, no centrifuge) Ideal Combination with XD s High Cell Densities 24

25 Simplify Downstream Process Direct Capture & Purification Adsorption Disposable pre-packed, Pre-sanitized columns Clarified, partially purified monoclonal DIRECT ELUTION Single use Bioreactor in XD mode Protein 25 A Mixed mode

26 EBA Experiments with XD very promising Results Feb-batch, classical Protein-A packed bed versus XD, EBA with Mixed Mode Ligand and Protein-A: Fed-batch, Filtration / Protein A Packed Bed XD, EBA - Mixed Mode Ligand Yield (%) Purity (%) HCP (ug/mg Mab) > 85 > (n=15) > 95 > (n=5) XD, PrA EBA >95 > (n=3) With high cell viabilities consistent t high h yield Purity still needs to be optimized (ligands/conditions) 26 HCP after column in normal range

27 If you have question for Rolf Douwenga send them to:

28