1. Case study: Challenges and learning in implementing ATF perfusion process
Dr. Jarno Robin
Principal Scientist
CMC_Biopharm upstream development
Novo Nordisk A/S, Hillerød, Denmark

2. Outline introduction to Novo Nordisk
case study: implementation of ATF perfusion process for improving yield prior to fase3 production
background & timeline
perfusion technology
contraints
new perfusion process (lab & production scales)
process challenges & stabilisation
conclusion:
yield improvement and quality
learnings

3. Novo Nordisk Corporate strategy
Novo Nordisk Way

4. Novo Nordisk around the world
R&D, production
Headquater,
Research Units,
production

5. Diabetes: A global pandemic

6. Part II
case study: implementation of perfusion process for improving yield prior to fase3 production
background & timeline
perfusion technology
contraints
new perfusion process (lab & production scales)
process challenges & stabilisation

7. Background & timeline
2008
Prod. Conc.
Feb : 3-days-Draw & fill (D&F) process approved for phase3 production.
very stable & robust process (cell growth & product quality)
BUT
very poor productivity
Issues regarding production capacity

8. Background & timeline
2008
Feb.: 3-days-Draw & fill (D&F) process approved for phase3 production.
Oct.: process challenge meeting: approval for attempting to change the production process from D&F to a perfusion process based on expectations to increase yield significantly (shaked flasks simulation).
Normalised yield
improvment (%)

9. Background & timeline
2008
Feb.: 3-days-Draw & fill (D&F) process approved for phase3 production.
Oct.: process challenge meeting: approval for attempting to change the production process from D&F to a perfusion process based on expectations to increase significantly yield.
Nov-Dec.: Finalising the ATF purchase for lab- & production- scales.
Jan-Jun.: Implementation of ATF-2 in lab scale and ATF-10 in production scale.
Apr-Jul: TDR meetings:
production medium chosen for phase3 production (3 upstream depts. involved).
perfusion process using the ATF chosen for phase3 production (3 upstream + 1 downstream depts. involved, coordination meeting).
Jul-Aug.: Master & production formulas in place for fase3 production planed from Oct (2 upstream depts. involved).
Q2/Q3: further process optimisation, MF/PF updated (2 upstream depts. involved).
Q4-2011/Q2-2012: issue regarding impact of complex raw-material quality on process performance (2 upstream depts. involved).
Q2/Q3: Process Justification & transfer to production support department.
2009
2011
2012

10. Why ATF ? : Perfusion devices available
Spin filter
Applisens BioSep
good in lab (intern filter)
 up-scaling: extern at large scale (temp., pH, O2 stress, pump & shear stress)
block at high cell conc.
cells in harvest
 good in lab
up-scaling, pump
5-10% cells in harvest
ATF (Alternating Tangencial Filtration)
Tangential Flow Filtration (TFF)
simple & upscalable (to 5m3)
no temp., pH, O2 stress
no pump & low shear stress
cell free harvest
reasonable cost
 possible product binding
good in lab
cell free harvest
 possible product binding
pump, up-scaling

11. ATF perfusion process at Novo Nordisk (Alternative Tangiencial Filtration)
Bleed
Medium
Harvest
Harvest
Patent no.: WO

12. Pictures of ATF (“Darth Vader” sword)

13. New ATF-process: constraints
Harvest stability of at least 2 days due to downstream process at large scale.
Short timeline for a complete process change (Q1/Q2-2009,~6 months)
Get a scalable perfusion process to the production facility using the ATF technology.
Get similar product quality as the material produced for tox and fase1 clinical trial using the D&F process .
Improve significantly the expression level (production capacity)

14. New process scalability from lab to production scale (2009)

15. Challenges: Complex raw-material quality impact on process performance at large scale

16. Soy hydrolysate has been analysed:
Challenges: Complex raw-material quality impact on process performance. Confirmation at lab scale
Soy hydrolysate has been analysed:
tracer element,
free amino acids
NIR

17. NIR analyses on HySoy batches
Work done by Erik Skibsted (ERSK)

18. Process stabilisation: focus on Zn concentration
w/ Zn spiking
w/o. Zn spiking

19. Conclusion: Yield improvment and quality
Process
Normalised Productivity
(%)
Quality parameter 1
Quality parameter 2
D&F
100
~95
0,03
ATF
250
90-95
0,06
600
~90
0,07

20. Conclusion: Learnings
targets achieved:
new process development & implementation within short timeline (6 months)
scalable perfusion process
significant increase in yield (up to 600%)
identical product quality between D&F and perfusion process
process stabilisation after unexpected significant impact of complexe raw-material on process performance
increased knowledge:
solving challenges
increased collaboration across lab & production scales
complex raw-material characterisation
know-how for next generation process and/or products for early implementation in the project plan:
Focus on correlation between cell line/medium/process

21. Aknowledgements: Dept.’s bioreactor team for dedicated technical work
Production & Research Unit, up- & down-stream for good collaboration
Vendors for support

22. Thank you !
for your attention