The document discusses the shift in downstream processing from a bottleneck to a driver of biomanufacturing. It notes that while high titers can reduce costs, downstream processing costs still dominate and present challenges to handle increased volumes and contaminant loads from improved processes. New technologies in filtration and chromatography using single-use disposables aim to improve throughput and capacity while reducing costs. The future of downstream processing involves revisiting past approaches with a focus on continuous improvement rather than identifying any single best solution.

1. “The new era of Downstream Processing:
From a Bottleneck to a Pacemaker”
Platzhalter Bild
BioManufacturing World Oct. 19 2010
Uwe Gottschalk VP Purification Technologies, Sartorius Stedim Biotech

2. Finally there seems to be a Bottleneck ...
6th Annual Survey of Biopharmaceutical Manufacturing.
Eric S. Langer, BioPlan Associates Inc.

3. Who is facing Limitations?
“Obviously there is no downstream bottleneck
if you have unlimited cash”
– K. John Morrow, Jr., PhD., 2009

4. Agenda
1.Improving throughput –
1.Improving throughput –
The Capacity Disconnect
The Capacity Disconnect
2.Process Economy – Cost
2.Process Economy – Cost
of Goods matter
of Goods matter
3.Discovering new Shores -
3.Discovering new Shores -
Fortune favors the Brave
Fortune favors the Brave
4.Benchmarks of the Future
4.Benchmarks of the Future
– No best, just better
– No best, just better
5.The Future of DSP –
5.The Future of DSP –
Revisiting the Past
Revisiting the Past
Data adapted from: F. Wurm Production of recombinant Protein Therapeutics
in Cultivated Mammalian Cells. Nature Biotechnology 22, 1-6 (2004)

5. Hot Topic: High Titer Processes
Improving Titres – MAbs (Bulk API)
Titre Improvements 4 Bioreactors
800
Important cost benefits as
titres go beyond 1g/L 700 Estimate of CoG based on
These diminish as we go 600
standard MAb process for
beyond 5g/L bulk drug substance
500
COG ($/g)
400
Beyond 5g/L 300
Downstream cost dominates
In this example the plateau 200
is just under $100/g 100
0
Challenge in DSP 0 2 4 6 8 10 12
Bioreactors decrease in size? Titre (g/L)
Cope with increased titres
2000L 5000L
Need to drive out costs
Implications for facility
design Results from Biopharm Services Generic MAb cost model
Bulk API Direct manufacturing costs
A universal cost model for single use systems in biomanufacturing.
Andrew Sinclair, BioPharm Services; Berlin Oct. 2007

7. Agenda
1.Improving throughput –
1.Improving throughput –
The Capacity Disconnect
The Capacity Disconnect
2.Process Economy – Cost of
2.Process Economy – Cost of
Goods matter
Goods matter
3.Discovering new Shores -
3.Discovering new Shores -
Fortune favors the Brave
Fortune favors the Brave
4. Benchmarks of the Future
4. Benchmarks of the Future
– No best, just better
– No best, just better
5.The Future of DSP –
5.The Future of DSP –
Revisiting the Past
Revisiting the Past

8. Challenges of a Modern Downstream Process
Technical challenge
 Sensitive and technically demanding products require processes with inherent
complexity and expensive infrastructure
 Need for robust & scalable processes for the entire DSP
 Increasing regulatory scrutiny (Comparability!)
Financial challenge
 Processes are fixed-cost driven (Investment vs Consumables)
 Manufacturing costs 15 - 25% of sales price
 Costs for DSP up to 75% of manufacturing costs
 Cost Balance Benefit for innovative treatments
 Biosimilars

9. Typical COG breakdown by
Category
MAb manufacturing: 6 x 2,000L tanks, 2g/L, 90% utilisation; 211 #/a; 527 kg/yr; invest 172 Mio Euro;
142 $/g; Sinclair 2006

10. Hot Topic: Use of Disposables

11. J. Zhou BPI Vienna 2008

12. Agenda
1.Improving throughput –
1.Improving throughput –
The Capacity Disconnect
The Capacity Disconnect
2.Process Economy – Cost
2.Process Economy – Cost
of Goods matter
of Goods matter
3.Discovering new Shores -
3.Discovering new Shores -
Fortune favors the Brave
Fortune favors the Brave
4.Benchmarks of the Future
4.Benchmarks of the Future
– No best, just better
– No best, just better
5.The Future of DSP –
5.The Future of DSP –
Revisiting the Past
Revisiting the Past

13. A Consensus – Value Chain in Bioseparation
High Titer Implications:
Increasing biomass and
contaminant levels
Protein A pool volumes
and step cost
DNA & HCP levels post
Capturing
Polishing load volumes
and conductivity
Pathogen clearance as a
moving target

14. Downstream Processing 1980
“If it ain’t broke, don’t fix it”
– Bert Lance, 1977

15. Downstream Processing 2010
“Le mieux est l’ennemi du bien”
(better is the enemy of good)
– Voltaire, 1772
“没有最好，只有更好”
(No best – only better)
– Chinese Movie Cliche

16. Agenda
1.Improving throughput –
1.Improving throughput –
The Capacity Disconnect
The Capacity Disconnect
2.Process Economy – Cost
2.Process Economy – Cost
of Goods matter
of Goods matter
3.Discovering new Shores -
3.Discovering new Shores -
Fortune favors the Brave
Fortune favors the Brave
4.Benchmarks of the Future
4.Benchmarks of the Future
– No best, just better
– No best, just better
5.The Future of DSP –
5.The Future of DSP –
Revisiting the Past
Revisiting the Past

17. Biomass Removal and Early Contaminant Clearance
addresses:
Increasing biomass and
contaminant levels
DNA & HCP levels post
Capturing
• New generation of lenticular
filtration media
• No Diatomeaceous Earth;
Synthetic
• Cell removal, clarification &
early on contaminant removal

18. The Vision of a Disposable Chromatography Process
Robert van Reis: Future Trends in Bioseparations; Recovery XII, 2006

19. Process Chromatography: What are the Options?
John Curling and Uwe Gottschalk
BioPharm Intl. October 2007
U. Gottschalk. Bioseparation in antibody manufacturing:
The good, the bad and the ugly.
Biotechnol Prog. 2008 May-Jun;24(3):496-503.

20. Medarex: Non-Protein A based Purification Processes: Scheme Evolution
2 columns + 1 column +
3 columns 1 membrane 2 columns 1 membrane
Unprocessed Bulk Unprocessed Bulk Unprocessed Bulk Unprocessed Bulk
CEX CEX CEX CEX
Chromatography Chromatography Chromatography Chromatography
Viral Inactivation Viral Inactivation Viral Inactivation Viral Inactivation
AEX AEX Membrane HCIC AEX Membrane
Chromatography chromatography Chromatography Chromatography
Mixed Mode Mixed Mode Viral Filtration Viral Filtration
chromatography chromatography
Viral Filtration Viral Filtration
Page 26
Alahari 2009

21. Two Birds – one Stone: Contaminant Precipitation at Pfizer and Medarex
addresses:
Fig 7a. precipitation based process Fig 7b. TFF based process
Contaminant
precipitation
Protein A pool volumes
TFF
and step cost
DNA & HCP levels post
HCP < 1000 ng/mg
CEX HCP < 10ng/mg CEX
Dilution
Capturing
Dilution
Q Membrane Q Membrane HCP < 1000 ng/mg
HCP BDL 2 g/ml
20 g/ml
Dilution
VF
Mix Mode
VF
Process Scale Precipitation of Precipitation of Process-Derived
Impurities in Mammalian Cell Culture Impurities in Non-Protein A
Broth; J. Glynn et al. In: Gottschalk U Purification Schemes for MAb; J. Wang
(ed) Process-scale Purification of et al. BioPharm Intl. 10/2009, 2-9
Antibodies. Wiley, NY.

22. Source: 2nd Annual Survey of the Bioprocessing Market
for Single-Use Solutions
Aspen Brook Consulting, 2010

23. Chromatography Technologies for DSP
Polishing (Membranes)
• Highly porous structure
• Pore size: 3 – 5μm
• Convective Flow
Capturing/IP (Resins)
• Minimal buffer use
• Bead size distribution: 15 -160 μm
• Average pore size: 15 - 40 nm
• Diffusion limited flow
• High capacity

24. Capture Costs: Why bother?
Jim Davis, Lonza Economics of Monoclonal Antibody Production: The relationship between upstream titer and
downstream costs; IBC San Diego March 2008

25. Protein A costs are not an issue at large scale with full total capacity utilization
6th Annual Survey of Biopharmaceutical Manufacturing.
Eric S. Langer, BioPlan Associates Inc.

26. Alternatives to Protein A Capture
• Product precipitation batch/continuous
• Impurity precipitation (followed by non-Protein A process)
• Alternative Capturing (Protein A Mimetics, Mixed Mode, CEX)
Issues: Selectivity, Scale up, Reproducibility, Comparability

27. addresses:
Protein A pool volumes
and step cost
D. Low BioManufacturing Paris 2007

28. Alternative Protein A Chromatography Formats:
Goal: Intensified Use/Volume Reduction
• Simulated Moving Bed (SMB) and related:
» Tarpon („single use flow path“)
» Novasep
» Chromacon
» Chromatan
» ...
______________________________________________________________________
• Expanded Bed Chromatography
» DSM/Upfront („single use flow path“)
Issues: Complexity, Scale up, Reproducibility, Comparability

29. Alternative Protein A Formats:
Goal: Low Cost – Real Single Use
2000: Oleosin Platform 2005: TMV Nanoparticles 2010: Bio Polyester Platform
Polyester
Synthase
Polyester Granule
Immunoabsorbent nanoparticles based on a 100-300 nm
tobacco mosaic virus displaying protein A
Limitation: Oleosin yields < 1kg/ha S. Werner et al. PNAS 103, 17678 - 17683
Grage, K. and Rehm, B.H.A. (2008)
Bioconj. Chemistry, 19(1):254-62.

30. Convective Media are Part of the Design Space in Polishing
...

31. Agenda
1.Improving throughput –
1.Improving throughput –
The Capacity Disconnect
The Capacity Disconnect
2.Process Economy – Cost
2.Process Economy – Cost
of Goods matter
of Goods matter
3.Discovering new Shores -
3.Discovering new Shores -
Fortune favors the Brave
Fortune favors the Brave
4.Benchmarks of the Future
4.Benchmarks of the Future
– No best, just better
– No best, just better
5.The Future of DSP –
5.The Future of DSP –
Revisiting the Past
Revisiting the Past

32. The Renaissance of Protein Purification
Michelangelo de Lodovico Buonarotti

33. Old Enabling Technology: Boring but Reliable
• Centrifugation
• Extraction
• Precipitation
• Filtration
• Crystallization
• UV-Inactivation

34. Thank you!
Uwe.Gottschalk@sartorius-
stedim.com