Scale-Up of a Continuous Purification Platform for Manufacture of Biologics Peter Levison, Ph.D. Executive Director Business Development, Pall Biotech, Portsmouth, UK 3rd International Symposium on Continuous Manufacturing of Pharmaceuticals: Implementation, Technology and Regulatory, London 3rd October 2018 This presentation is the work product of Pall Corporation and no portion of this presentation may be copied, published, performed, or redistributed without the express written authority of a Pall corporate officer. Š 2018 Pall Corporation.
Pall Continuous Bioprocessing Solution from PD to Manufacturing PD to Manufacturing
Cell Culture Bioreactors
Clarification Cadence Acoustic Separator Cadence™ Acoustic Separator
Depth + 0.2 µm
Cadence Inline Concentrator (ILC)
Purification Capture – Cadence BioSMB + Cadence Virus Inactivation
2
Depth + 0.2 µm
Polish – Cadence BioSMB
Formulation Virus Filtration
Cadence ILC
Cadence Inline Diafiltration (ILDF)
0.2 µm
Large Scale Cadence Acoustic Separator Product Options
100 LPH Add-On 50 LPH Base Skid
3
350 LPH
Total Cell Density (x106 cells/mL)
Cadence Acoustic Separator – Scale Up Feed TCD
30 25
Stage 1
20 Stage 2
15 10
Stage 3
5
Stage 4
0 50 L/h
4.2 L/h Scale
50 L/h Scale
4.2 L/h Scale
Feed
Perm
% Red
Feed
Perm
% Red
TCD (106 cells/mL) Turbidity (ntu)
27.9 1496
2.1 251
92 83
31.6 1385
2.5 158
92 89
Viability (%)
81.5
n/r
81.5
n/r
Vol. Yield (%)
89
90 4
Scalable Continuous Chromatography
Process Development
Full Scale Cadence BioSMB Process 80 and 350
Cadence BioSMB PD Attribute
Patented Valve Cassette Design
Cadence BioSMB Process 80 Designed for perfusion applications with a single use flow path that can be set up in 30 minutes or less
Production time of 6 h for a 2000 L bioreactor with a single use flow path that can be set up in 30 minutes or less
3 – 16 *
3–8
3–8
0.6 – 3 L/h
10 – 80 L/h
30 – 350 L/h
Modular design Flexibility in Column arrangement (series or parallel) Number of Columns Flow Rates
Cadence BioSMB Process 350
Only multicolumn chromatography system that makes 24-hour process development a reality
* for ease of integration development 5
Cadence BioSMB Process 350 – Conditions Feed material: 400 L clarified cell supernatant with 5.8 gm/L mAb Chromatography sorbent: KANEKA KanCapA♦ PD Scale
Process Scale
4.0 mL/min
37 L/hr
11.2 mm ID x 5 cm H
14 cm ID x 5 cm H
Column volume
5 x 5 mL = 25 mL
5 x 0.77 L = 3.85 L
Residence time
3:04
3:04
5
5
46.4 gm/L
46.4 gm/L
Flow rate Column dimensions
Number of columns Operating binding capacity
> 150 x Scale-up 400 L processed in 13 cycles over 11 hours Data presented by Mark Brower (Merck, Kenilworth NJ) at Bioprocess International Conference, Boston, 2016 6
♦ KANEKA KanCapA is a trademark of KANEKA Corporation
Process Consistency 13 cycles of operation Consistent UV elution performance Consistent impurity clearance
Data presented by Mark Brower (Merck, Kenilworth NJ) at Bioprocess International Conference, Boston, 2016 7
Performance Comparison Batch Reference
BioSMB PD
BioSMB Process 350
Product conc.
9.54 gm/L
13.84 gm/L
13.85 gm/L
Product yield
98%
97%
97%
Aggregate
0.45%
0.72%
0.65%
LRV DNA
n.a.
4.2
5.0
LRV HCP
2.4
2.6
2.5
16 gm/L/hr
56 gm/L/hr
56 gm/L/hr
Spec.Prod. Comparable process Cycles
3
13
Column diameter
40 cm
14 cm
Column height
16 cm
5 cm
20 L
3.85 L
Protein A volume
Data presented by Mark Brower (Merck, Kenilworth NJ) at Bioprocess International Conference, Boston, 2016 8
Cadence BioSMB Process 350 System – User Data / Feedback A total of 2.3 Kg mAb (400 L at 5.8 g/L) was purified in 12 hours (13 cycles) Only 3.85 L of Protein A media was required (KANEKA KanCapA) A comparable batch process would require 20 L of Protein A media Five columns was the optimal configuration This is approximately 10% of the maximum capacity of the
Cadence BioSMB Process system Batch process was translated to continuous at PD scale and
transferred to Process Scale (150x) in less than 3 weeks Operation (HMI) of Cadence BioSMB Process system is
relatively easy (comparable with Cadence BioSMB PD) Result for Process Scale and PD Scale are very similar
(performance and product quality attributes) The Cadence BioSMB PD process successfully scaled up >150-fold 9
Data presented by Mark Brower (Merck, Kenilworth NJ) at Bioprocess International Conference, Boston, 2016
Cadence BioSMB Process 80 System – User Data / Feedback mAb process was translated from batch to continuous at PD scale and
transferred to Process Scale (40x) using a 4 column system
Productivity [g/(L*d)]
1000
800
600
400
200
0
Setting 1
Setting 2 Batch
BioSMB PD
Setting 3
BioSMB Process 80 Data presented by Ozan Ötes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018
10
Cadence BioSMB Process 80 System – User Data / Feedback A modified 3 column process was operated continuously for 10 days CQA’s remained consistent throughout the study Monomer content by SEC 100.0
Purity [%]
99.0 98.0 97.0 96.0 95.0 0
2
4
6
8
Day of experiment
11
10
Data presented by Ozan Ötes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018
Cadence BioSMB Process 80 System – User Data / Feedback A modified 3 column process was operated continuously for 10 days CQA’s remained consistent throughout the study 200
HCP [ppm] Res. Protein A [ppm]
Impurities [ppm]
160
120
80
40
0 0
2
4
6
8
Day of experiment
12
10
Data presented by Ozan Ötes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018
Cadence BioSMB Process 80 System – User Data / Feedback Consistent operation was reported across multiple cycles over 10 days UV Absorption [AU]
2.50 Cycle 01 Cycle 02 Cycle 03 Cycle 04
2.00 1.50
Cycle 05 Cycle 06 Cycle 07 Cycle 08 Cycle 09
1.00 0.50 0.00
Time in Cycle [hh:mm] 120.0
Conductivity [mS/cm]
Cycle 01
100.0
Cycle 02 Cycle 03
80.0
Cycle 04 Cycle 05
60.0
Cycle 06
40.0
Cycle 07 Cycle 08
20.0
Cycle 09
0.0
Data presented by Ozan Ötes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018
Time in Cycle [hh:mm]
13
Cadence BioSMB Process 80 System – User Data / Feedback Opportunities for cost reductions and process improvements
Save Time
Save Resin
Develop more projects
CTM
Lower ROI
More batches per year
Production
Higher throughput
Lower COGs
14
Faster Processes
Data presented by Ozan Ă–tes (Sanofi, Frankfurt, Germany) at BioProcess International European Summit, Amsterdam, 2018
Cadence Virus Inactivation System Comprised of one control unit and one or
two mixers Designed for both continuous and
batch operations Fully automated semi-continuous operation – For processing up to 2000 L fermentation volume – Single-use gamma irradiated flow path – GMP ready – Uses proven low pH virus inactivation methodology – Capacity up to 30 L/h volume
15
Cadence Virus Inactivation Operation Concept
16
Single-Use System Error free installation ensured by – Tagged connections – Shadow boarding – Unique genders and sizes on certain connections to make wrong assembly impossible
Mixer and manifolds can be installed in under
30 minutes Sample port in recirculation loop
for highly representative sampling No sample port on mixer
biocontainer
17
Cadence Virus Inactivation System
Phi 6 inactivation Start titer 1.33E+07 Cycle 1 2.00E+07 Cycle 2 NA Bag wash post cycle2 Bag control (no titration) 3.40E+07
End titer 0 0 0 3.30E+07
Integrated with Cadence BioSMB Process System, or standalone Single use flow path with installation in less than 30 minutes Capable of fully automated processing for batch chromatography No dead legs – Tested with: – Bacteriophage – B. diminuta – Riboflavin
Fully automated, designed for easy integration into continuous processes up to 2000L cell culture volume 18
Validation Results Test data available for testing with: – Bacteriophage (used as a virus analogue) to show effectiveness of the inactivation methodology – Bacterial challenge with Brevundimonas diminuta (B. dim) to show effectiveness of the design in eliminating hold-ups and carry over The bacteriophage data showed: – No active bacteriophages in the inactivation mixer and recirculation loop after completing two cycles, demonstrating the efficiency of the dosing, mixing and recirculation methodology – Controls showed no damage to the protein from pumping and mixing The B. dim testing data showed: – Complete washout of the target organism to below the limits of detection, indicating that the system does not have any detectable dead legs
19
Cadence BioSMB and Cadence Virus Inactivation Systems Together– Designed to be Integrated as a Combined Unit Operation Both systems can be integrated to run
continuously Automation – Connected directly or via control system to upstream and downstream operations
Process – Connected to Cadence BioSMB product port by interconnection manifold – Ability to collect whole elution peaks based on weight change in mixer or opening / closing of product valve
20
Evolution in Bioprocessing Key driver is time to market
Single-Use Technologies
Stainless Steel
Modular Construction
Continuous BioProcessing
First Movers
General acceptance
Second tier
Aging Technologies Adapted from: Morten Munk, PDA Annual Meeting 2016
Impact of Continuous BioProcessing Improved Facility efficiency Manufacturing flexibility
Reduced Processing time Operating costs Footprint Capital outlay 21
Detailed Commercial CoGs Breakdown: 20 batches / year at 6000 L and 5 g/L 16 Cost per Gram ($/g)
14 12 10 8
Other Labour Consumables Materials Capital
6 4 2 0 SS Batch
SU ICB SS Batch Platform Batch
Clarification
SU ICB SS Batch Platform Batch
Purification
SU ICB SS Batch Platform Batch
Polishing
SU ICB Batch Platform
Final Form.
ICB Platform exhibits 30% savings over SS Batch and 16% over SU Batch – $4.7M and $2.2M annual savings, respectively 22
Thank You peter_levison@pall.com