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Webinar summary:
1000L-scale implementation of fully connected, disposable, advanced DSP platform for next generation mAb production.
Within the biopharmaceutical industry, there is a significant shift toward higher productivity processes resulting in improved economics without compromising robustness. Therefore, integrated continuous production technologies are of greatest interest.
Next Generation Biopharmaceutical Downstream Process is a European-funded collaborative project that aims at implementing a fully integrated manufacturing platform for biosimilar mAb based on continuous chromatography, in combination with single-use disposable technologies for all unit operations of DSP on pilot/small production scale together with incorporation of advanced analytical tools.
In this webinar, you will see:
* new DSP purification template producing > 3.3 kg of mAb in 2.5 days in less than 30m²
* proof of concept for the mAb manufacturing of tomorrow
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Implementing a Fully Single-Use, Integrated mAb Biosimilars Purification Platform for Next Generation Manufacturing
1. Merck KGaA
Darmstadt, Germany
Implementing a fully single-use,
integrated mAb biosimilars
purification platform for next
generation manufacturing
Josselyn Haas Durr
European Biomanufacturing
Engineer Manager
2. 2
The life science business of
Merck KGaA, Darmstadt, Germany
operates as MilliporeSigma
in the U.S. and Canada.
5. Title
Next-generation biopharmaceutical downstream process
Acronym: nextBioPharmDSP
Webpage: www.nextbiopharmdsp.eu
General 4 year project (March 1, 2015 – February 28, 2019)
Partners
Industry: Lek Pharmaceuticals d.d. (Slovenia), Sandoz GmbH
(Austria), Millipore SAS (France)
Academia: University of Natural Resources and Applied Life
Sciences (Austria), Karlsruhe Institute of Technology (Germany),
National Institute of Chemistry (Slovenia)
SME: National Systems srl (Italy)
Objective
Develop and implement a more efficient, cost-effective and
environmentally friendly downstream process for the
manufacture of monoclonal antibodies and biosimilars.
Horizon 2020: EU-funded Program for Research & Innovation
Snapshot
5
Budget
Total budget: 10,6 mio €
EU funding: 8,4 mio €
70 % for industry
100 % for academia
6. 1000L Fed-Batch
Bioreactor
Optimizing Biopharmaceutical Downstream Processing
Establish fully connected disposable DSP platform for biologics
production
Bioreactor Centrifuge Filtration Protein A VI CEX b/e AEX f/t VF UF/DF
Bioreactor +
pre-treatment
Filtration Continuous
VI
Continuous
capture
Carbon & AEX
f/t
CEX & VF
f/t
UF/DF
Continuous DSP - 3 days - 4kg
Capture Intermediate / Polishing VF UF/DF
Primary
Separation
6
Batch
Process
Continuous
Stainless
Steel
Format
Single-Use
10. Surface installed 15m² 3,85m²
Clarification train
Two-step clarification:
D0HC X0HC
One-step clarification:
pDADMAC 40 MS
Consumable cost Equivalent (including Flexware® assemblies and flocculant)
Water for flush 1500 L 385 L
Hold up volume 150 L 100 L
Operation time 1,5 day 1 day
pDADMAC pre-treatment followed by Clarisolve® 40MS depth filter
exhibit benefits compared to traditional set up
5–10M TC/mL
72–98% Viability
1040–2600 NTU
harvest turbidity
50% safety factor
€
3 mAb
1000L CHO
- 75%
- 75%
- 33%
- 33%
2:1
=
10
Primary clarification
11. Primary clarification
Large scale implementation
11
Bioreactor Flocculation
in the
bioreactor
pDADMAC
10 % stock
solution
Water
Buffer
Bioburden
reduction
Clarisolve®
40 MS depth filter
Clarified
harvest
14. Small beads offer better performances for continuous capture
Particle size: 35 – 49 μm
Particle size: ~ 85 μm
Particle size: 35 – 49 μm
Particle size: ~ 85 μm
Batch chromatography
Continuous chromatography
10 Protein A resins evaluated
• DBC for several residence times (RT)
• BT curve shape & BT point
• Removal of HCP, aggregates and step
yield no significant differences
• DNA removal some differences - not
critical
14
Protein A resins screening
®
®
Resin
Resin
15. Continuous Chromatography
Continuous capture on 3 columns: a robust and consistent operation
15
Processes developed on several resins – optimized step lengths
50 – 60 cycles performed – consistent performance
Method duration: 65 – 75 min / cycle
RT 1.5 min, high loadings (60 – 70 mg/mL)
High productivity
and resin utilization
16. Development of a disposable 3-columns system
• Fully disposable flowpaths (incl. flowmeters, pump
heads and UV)
• 8 to 120 l/h per line
• 30 days of continuous operation
• Fit for columns from 7 to 25 cm i.d.
• 3 inlets product line, 6 inlets buffer line
16
Continuous Multi-Column (CMC) Capture
17. From concept to reality
17
Continuous Multi-Column (CMC) Capture
19. Viral inactivation is dependant on pH, buffer type and temperature, but NOT
on mAB concentration
19
Virus inactivation at low pH is rapid and requires less than
5 minutes.
The kinetics depend on pH, temperature, and buffer
conditions, and are not dependent on protein
concentration.
The incubation chamber that can achieve a target
inactivation time with an appropriate safety factor must
be designed appropriately.
21. From 3D concept to reality
21
Semi-continuous virus inactivation
Pumps for pH
regulation
C2
WYE inlet Flexware®
and F1/F2 valve
C1 Acid/Base bags holder
C1 to C1B
pump
C1B
23. pH 5
Protein A elution material after VI
AEX FT-CEXCarbon
AEX
FT-CEX
FT-CEX
pH 7
MAb
acidic pI basic
LowMWhigh
Larger acidic
HCP, DNA, and viruses
Anion Exchangers
HCP
Cation Exchangers
Smaller HCP’s
and cell culture components
Activated Carbon Functionality Media
23
Toolbox Approach
Flow-Through Polishing
25. Millistak+® CR40 depth filter is the main contributor to HCP reduction
level
25
Activated Carbon
Binding: van der Waals
interactions
Size-based selectivity
Protects AEX
CAP.E. AFTER AC AFTER AEC AFTER CECCAP.E. AFTER AC AFTER AEC AFTER CEC
26. Eshmuno® CP-FT resin outperforms CEX membrane adsorbers for
aggregates removal in Frontal chromatography mode
26
FT cation exchange
27. Unit Operation Technology Selection
Flow-Through Polishing – Implementation Strategy for 1000L Reactor
Protein A
Eluate after
VI
Millistak+®
CR40
Activated
Carbon Depth
Filter
(1.2 m2)
Eshmuno®
Q
AEX FT Resin
(2 x 1.1L)
Eshmuno®
CP-FT
CEX FT Resin
(2 x 1.1L)
Viresolve®
Pro
Virus Filtration
(0.22 m2)
adjust pH=5
pH = 7
Conductivity < 5 ms/cm
27
30. DSP continuous process control: system handshaking
Bioreactor +
pre-treatment
Clarification
Continuous
Virus
Inactivation
Continuous
capture
f/t Carbon
f/t AEX
f/t CEX
& VF
UF/DF
pH
adjustment
Switch of VIN
collection tank
Full VIN tanks stop CAP
VIN ready starts
Carbon/AEX
pH adj. starts
CEX/VF
Breakthrough
detection
Peak
detection
Flow
VPE
VIN = Viral Inactivation30
31. Continuous capture during the 4 runs
Data Run 1 Run 2 Run 3 Run 4
Number
of cycles
Total 90 92 94 139
col 1 30 30 32 46
col 2 30 31 31 46
col 3 30 31 31 47
Loading speed
(cm)
180 240 210 210
Elution speed
(cm)
180 280 280 280
31
32. 32
Capture – CMC skid
Trends (run 4) demonstrate the consistency of the capture process
32
33. 4 technical runs
at 1000L scale
2,5 kg mAb purified in 2,5 days
> 80% yield all runs and steps considered
9–12 g/l mAb concentration after Viresolve® Pro solution
CONSISTENT purity and quality over runs
• No bioburden
• RPC and CZE profiles comparable to reference33
38. Main benefits
38
€
• Facility investment: 35%
• Running costs: 30%
• Cost of materials: 50%
• CO2 emissions: 25%
• Reduced water, buffer, resins consumption (water 10x)
Productivity
Footprint
Economics
Single-Use
Environmental
• Reduce bioburden risk
• No carry-over issues
• No cleaning, regeneration, steaming
• Rapid change-over
• Complete DSP in 30 m2 (10-15x smaller)
• Elimination of large intermediate hold tanks
• Flexibility (mobile equipment)
• Capable of processing up to 3000L harvest in 24 hours (up 12 kg of mAb)
nextBioPharmDSP project helps to make highly efficient
biopharmaceuticals more accessible to patient
39. You are invited!
Next Generation Bioprocessing
Forum
October 8-9, 2019
M Lab™ Collaboration Center
Molsheim, France
40. Trademarks and logos used in this presentation are the property of companies
collaborating in the nextBioPharmDSP project.
Thank you!
www.nextbiopharmdsp.eu
This project has received funding from the European
Union’s Horizon 2020 research and innovation
program under grant agreement No 635557