This document discusses the development of large-scale mammalian cell culture facilities from the 1980s to present day. Early facilities in the 1980s utilized simple semi-manual systems with basic automation and validation. Facilities evolved in the 1990s to reduce complexity and costs through simplifying stainless steel bioreactors and adopting single-use disposables. Present day facilities since 2008 widely accept large-scale disposable bioreactors up to 2000L in volume. The next generation of facilities aims to further reduce footprint and costs through integrated single-use systems for media preparation, buffer storage, and harvesting.

1. Large scale Mammalian plant design from 1980’s to present day Dave WoltonVP manufacturing1

2. OverviewCMC

3. Development of Large scale Mammalian Monoclonal factories

4. 1985-1990

5. 1990-1995

6. 1995-2006

7. Present day

8. Next Generation Factories

9. Impact of the new design on the rest of the organisation

10. CMC prototyping of disposable harvest systems

11. Summary2

12. CMC Biologics – Overview Target & Lead ID Pre-toxToxPhase I Phase II Phase IIICommercial ProductionCMC BiologicsContract Manufacturer of Biological Therapeutics utilizing MAMMALIAN & MICROBIAL CELL CULTURE TECHNOLOGIESCell-line Development – CHEF1™ technology

13. Upstream/Downstream – process development and manufacture

14. Analysis, Characterization, Formulation, Quality and Regulatory3

15. Development of Large scale Mammalian Monoclonal factories Late 80’s4

16. Late 80’sThe dawn of automationSimple semi manual fermentorsValidation becoming more involvedContainment being actively pursuedCIP/SIP systems becoming more complicatedAutomation seen as reducing Headcount, Turn-round timeOperator errors5

17. Early 90’s6

18. Early 90’sSome companies begin to realise complex automation systems have drawbacks. They see increasing HeadcountComplexity of plantTurn-round timesComplexity of investigations (variables)Facility build costsFacility construction times7

19. Early 90’sExample of early 90’s BioreactorAdditive manifoldAir OutAir to headspaceAir to spargerJacket heat exchangerSample device8

20. Late 90’s to 20069

21. Late 90’s to 2006Some companies start to reduce cost by simplifying.Stainless steel reactors are simplified by reducing numbers of valves/items.Disposables reduce the need for CIP/SIP. InnovationsTubing weldersLynx valvesPod filtersWave reactorsSUM’s10

22. The fastest CIP? Reduce the number of inlets that need cleaning.How to simplify stainless vessels11

23. How to simplify stainless vesselsAdditive manifolds12

24. How to simplify stainless vesselsAir Out13

25. How to simplify stainless vesselsAir To Head Space14

26. How to simplify stainless vesselsAir To Sparger15

27. How to simplify stainless vessels16Number of sterile boundary elastomersPre Lynx ModificationsApproximately 70Post Lynx modificationsApproximately 30

28. Present Day17

29. Single use bioreactors18

30. Present dayElan’s plant design breaks new ground (2008).Large scale disposables are widely accepted2000L reactors are launched onto the market (2010)19

31. Next generation factories20

32. The Facility – the next generation of plant after élan2g/l monoclonal antibody- fed batch10 x 4,000L batches per month80 cm column, 3 cycles = 2 x 12,000L stainless plant21

33. The Facility – The next generation after ÉlanFoot print ComparisonPurificationBioreactors.XXBuffer prep.Media prep.22

34. The Facility – the next generation of plant after élanHow do you make media next to the bioreactor without contaminating the room with powder?23

35. Contained media mixing – GEA24

36. Contained media mixing – HycloneHyclonePowdertainer25

37. Contained media mixing - Lormac26

38. Use of Buffer towers in Downstream worked example27

39. The Facility – the next generation of plant after élanHow do you make and store up to 6,000L of buffer and not use large numbers of buffer bags?28

40. Buffer towers - PrototypeAnswerBy positioning 2,000L buffer towers close to the column and ‘topping up’ from 1,000L single use mixers.Prototype29

41. Buffer towers - worked exampleBuffer ModelingBuffer storage in 1000L Buffer hold bags VS Five 2,000L buffer towers; replaced monthlyBuffer b Adsorption chromatography buffer - Max 4,000LBuffer a UltrafiltrationDiafiltration buffer – Max 6,000L30

42. The Facility – the next generation of plant after élanBuffer bBuffer a31