Membrane Chromatography Solutions for Single-Use, Intensified mAb Purification

The life science business of Merck KGaA,
Darmstadt, Germany operates as
MilliporeSigma in the U.S. and Canada.
Membrane Chromatography
Solutions for Single-Use,
Intensified mAb Purification
Gary Skarja, Ph.D., Head of Membrane Chromatography R&D
Mochao Zhao, Global Product Manager, Single-use Membrane
Chromatography

The life science business
of Merck KGaA, Darmstadt,
Germany operates as
MilliporeSigma in the U.S.
and Canada

Agenda
1
2
3
Cost Considerations for Mab Purification: Opportunities for
Improvement
Natrix® Technology
Natrix® A: Intensified MAb capture
4 Towards A Fully Single-Use Chromatography
Platform

Cost Considerations for
Mab Purification:
Opportunities for
Improvement

Cost Considerations at Different Development Phases
Membrane Chromatography Solutions for Single-Use, Intensified mAb Purification5
Monoclonal Antibody Purification
Clinical Production
Fail Cheaper Fail Faster
• Delay initial capital
investment when risk is high
• Reduce investment that
requires amortization to be
cost efficient
• Accelerate process
development
• Reduce construction lead
time (if new facility or plant
retrofit is required)
• Reduce non value-adding activities
• Improve process productivity
References:
1Biotechnology Innovation Organization, Biomedtracker, AMPLION. Clinical Development Success Rates 2006-2015;
2016.
2Deloitte Touche Tohmatsu. Therapeutic Monoclonal Antibodies: Insights, Strategies and Data – How US Biotechnology
Companies Pioneered the Path to Clinical, Regulatory and Market Success for Monoclonal Antibodies Recap; 2011.
3Joseph A. DiMasi, Henry G. Grabowski, Ronald W. Hansen. Innovation in the Pharmaceutical Industry: New Estimates
of R&D Costs. Journal of Health Economics. Volume 47 (20-33); 2016

Cost Considerations at Different Development Phases
Monoclonal Antibody Purification
Transition to Commercial Production
Facility Investment Facility Utilization Scale of Operation
Reduce footprint
• Downsize unit operation and supporting
hardware
• Simplify process architecture
• Reduce areas associated with non value-
adding activities
• Beyond capital expenditures: plant
utilities, maintenance and insurance
Improve flexibility and modularity
• Capacity shortage
→ Increase production rate
→ Scale Up versus Scale Out
• Over capacity
→ Possibility of multi-product
implementation
→ Reconfigure suite for new processes
Identify dominate cost categories for
targeted production scale
• Ex. Micro-facility vs Low-volume Multi-
product vs Large-volume Single-product
Evaluate trade-offs between major
cost categories
• Ex. Increase in “Consumable Cost” due
to SUT implementation is often
compensated by decrease in “Capital,
Labor, Maintenance Cost”
Manufacturing Process Drives Facility
• Each unit is not a standalone operation → Advancement in one unit can unlock improvements in the overall process
architecture
• Economic improvement requires holistic considerations at PD phase

Plug-and-Play, Single-Use per Batch Format
Membrane Chromatography Solutions for Single-Use, Intensified mAb Purification
Membrane Chromatography Solution
❖ Reduce non value-adding activities and associated capital, labor
& material expenditures
• Column packing/unpacking
• Cleaning, storage (in and out), and associated validations
• Column hardware maintenance and service
❖ Flexibility enables rapid changeover to improve facility utilization
❖ Eliminate cross-contamination risk
❖ Reduced bioburden concern since storage is no longer required
❖ Shift Fixed Cost to Variable Cost
7
AEX
Resin
AEX
Membrane

High Productivity Enables Unit Right-Sizing
8
=
𝑔 (𝑚𝑎𝑠𝑠 𝑜𝑓 𝑟𝑒𝑐𝑜𝑣𝑒𝑟𝑒𝑑 𝑚𝐴𝑏)
ℎ𝑜𝑢𝑟 (𝑡𝑜𝑡𝑎𝑙 𝑝𝑟𝑜𝑐𝑒𝑠𝑠 𝑡𝑖𝑚𝑒) × 𝐿 (𝑚𝑒𝑑𝑖𝑎 𝑣𝑜𝑙𝑢𝑚𝑒)
=
𝑟𝑒𝑐𝑜𝑣𝑒𝑟𝑦 × 𝑙𝑜𝑎𝑑 𝑐𝑎𝑝𝑎𝑐𝑖𝑡𝑦 × 𝐿 𝑚𝑒𝑑𝑖𝑎 𝑣𝑜𝑙𝑢𝑚𝑒 × # 𝑜𝑓 𝑐𝑦𝑐𝑙𝑒𝑠
𝑐𝑦𝑐𝑙𝑒 𝑡𝑖𝑚𝑒 × # 𝑜𝑓 𝑐𝑦𝑐𝑙𝑒𝑠 × 𝐿 (𝑚𝑒𝑑𝑖𝑎 𝑣𝑜𝑙𝑢𝑚𝑒)
=
𝑟𝑒𝑐𝑜𝑣𝑒𝑟𝑦 × 𝑙𝑜𝑎𝑑 𝑐𝑎𝑝𝑎𝑐𝑖𝑡𝑦
𝑐𝑦𝑐𝑙𝑒 𝑡𝑖𝑚𝑒
Chromatography
Productivity
Evolution in resin binding
capacity expected to only
marginally improve
productivity
Dominated by residence time requirement
Opportunity for significant productivity
improvement
• Residence time decrease from minutes to
seconds
• Process time from hours to minutes

High Productivity Enables Unit Right-Sizing
Low-productivity resin chromatography operation requires over-sizing to balance
between process time and product quantity requirements
• Large footprint for unit and supporting hardware
• Requires amortization over large number of batches to be cost efficient
9
High-productivity membrane chromatography allows right-sizing based on
capacity need, and full media utilization within a batch
• Downsize unit and footprint with no “wasted” capacity – delay expenditure
during high-risk clinical development phase
• Cost efficient without amortization

Natrix® Technology
Advances in Membrane Sciences
Flexible, reinforcing fiber mesh provides
strength and structure
Reinforcing mesh is filled with functionalized,
porous gel
✓ Gel provides binding groups and final pore
structure
Functionalized, durable composite membrane
is created in a single step
Identical functional binding group chemistry
as resins – C, Q , mixed-mode, affinity
11

NATRIX® MEMBRANE
Bead-like binding with >20x
shorter residence times
Unique 3D porous structure maximizes
ligand density & eliminates diffusive
transport speed limitations
+ Bind & Elute cycle times in minutes
+ High binding capacity
➔ Maximized Productivity
MEMBRANES
+ Short Residence Time
-Low Binding
RESIN BEADS
+ High Binding
- Long Residence Time
Combining the Best of Resin and Traditional Membrane Adsorber
Performance Features
12

Enabling A Fully Single-Use Chromatography Platform
Plug-and-Play, High Productivity Chromatography Membrane
13
Plug-and-Play devices
• eliminates column packing &
qualifications
Single-Use capsules
• eliminates column cleaning/
storage and associated
validations
• eliminates risk of cross
contamination
40x higher flow rate
than bead resins decreases
process time

Natrix® A:
Intensified mAB capture

Key Considerations
Natrix® A Membrane Design & Development
Membrane Matrix Design Pro A Ligand Selection
Pro A Membrane
Property Optimization
✓ Low synthetic complexity &
cost
✓ Suitable membrane reactive
group chemistry
✓ Non-binding matrix
properties
✓ Screened several promising
engineered Pro A ligand
candidates
✓ Selected best combination
of dynamic binding capacity
and alkaline stability
✓ Pro A Membrane binding
capacity & permeability was
varied by modifying
reaction formulation and
conditions
15

MAb Capacity & Impurity Clearance
Natrix® A Membrane Performance
Natrix® A membrane shows good dynamic
binding capacity over a range of very short
residence times
Enables very high productivity mAb capture
processes and low-footprint device designs
Natrix® A membrane HCP clearance is similar to
or better than Protein A resins – demonstrated
over a wide range of mAb feeds
16

333,500
312,000
272,000
189,000
176,000
131,000
361,000
SHC + SPTFF + SHRAA Precip +
Clarisolve + SHR
pDADMAC +
Clarisolve + SHR
SHC + SPTFF + X0HC
+ SHR
pDADMAC +
Clarisolve + X0HC +
SHR
X0HC + SHRFEED
974
462
214
1119274
1,466
MAb Capacity & Impurity Clearance
High performance clarification could enhance capture performance
→ Enabling further process simplification
Clarified Harvest HCP (ppm)
Natrix® A Eluate HCP (ppm)
17

Bind & Elute Cycling Consistency
Natrix® A membrane has been optimized to exhibit consistent
impurity clearance over 100+ cycles operating at short
residence time (seconds)
• Consistent purification & elution performance over 100
cycles with 0.1 M NaOH CIP each cycle
• High productivity media enables reduced device size
and/or process time
• Media life fully utilized in a single batch, simplifying &
improving the economics of the downstream process
18

Right-Sized, Single-Use per Batch Capture To Fully Utilize Media Life
High Productivity Enables Rapid Multi-Cycling Operation
Rapid Multi-Cycling with Natrix® A
19

Evaluation of Natrix® A screening device using “typical” capture conditions BUT
at very short membrane residence time
• Natrix® A residence time: 6 seconds
• Typical resin residence times: 2 – 6 minutes
Include caustic cleaning step in every cycle to confirm stability of performance
over 100 cycles
Non-optimized buffer condition
Rapid Bind & Elute Multi-Cycling Operation
Natrix® A Screening Device
Step Buffer
Equilibration Neutral pH, low conductivity
Load Clarified harvest
Wash 1 Neutral pH, low conductivity
Wash 2 Neutral pH, higher conductivity
Wash 3 Neutral pH, low conductivity
Elution Low pH acetate
Strip Acetic acid
Cleaning 0.1 N NaOH
Re-equilibration Neutral pH, low conductivity
20

Consistent High Performance with Rapid Multi-Cycling Operation
Natrix® A screening device shows
very consistent chromatographic
performance over 100 bind & elute
cycles at 6 second residence time
Chromatogram overlay of every
10th cycle for 100 bind & elute
cycles with clarified mAb feed
21

Consistent High Performance with Rapid Multi-Cycling Operation
Natrix® A screening device shows
very consistent chromatographic
performance over 100 bind & elute
cycles at 6 second residence time
Chromatogram overlay of every
10th cycle for 100 bind & elute
cycles with clarified mAb feed
22
Elution properties are consistent over 100 bind & elute cycles,
supporting rapid multi-cycling operation
❖ Good mAb yield
❖ Excellent impurity clearance
❖ Low HMW content

Towards A Fully Single-Use
Chromatography
Platform

Aggregate and Impurity Removal
High Throughput Flow-Through Polishing Tools
24
Natrix® Sb (In Development)
• Salt tolerant strong cation exchange with HIC modality
Efficient aggregate & HCP removal at high mAb
loading in flow-through mode enables high
productivity polishing, providing the following
benefits:
✓ right-sized purification step
✓ reduced equipment footprint
✓ unlocks true single-use operation

Viral Clearance and Impurity Removal
High Throughput Flow-Through Polishing Tools
NatriFlo® HD-Q (Commercial)
• Salt tolerant strong anion exchange media
25
✓ Impurity clearance is independent of the
load up to 20 kg/L
✓ Good purification performance at exceptional
loading capacity enables downsizing of the
unit operation
✓ Good viral clearance over a range of buffer
and pH conditions enables a wide design
space for operation

Natrix® A
Natrix® Sb
NatriFlo®
HD-Q
Proof of Concept
Towards A Fully Single-Use Chromatography Platform
26
HCP HMW Yield
247 ppm 10.4% 90%
47 ppm 0.5% 93%
5 ppm 0.5% 95%
mAb 8: 142kDa; pI 8.2
Feed HCP: 342,188 ppm

Operate More Simply and Effectively, From Lab Bench to Commercial
Towards A Fully Single-Use Chromatography Platform
Achieve Purity Target
Accelerate Innovation
• Speed up process development
• Reduce validation requirements during clinical development
• Reduce construction lead time thanks to smaller footprint
and less requirement on supporting hardware/area
Reduce Quality Risk
• Eliminate risk of cross
contamination between batches
• No storage required – less
bioburden concerns
Improve Facility Utilization
• Greater flexibility enables more rapid production changeover
• More batches annually
• Multi-product facilities
• Rapid response to varying market demand
• Shorter time to reconfigure suite for new processes
Mitigate Clinical Risks
• Delay large expenditures
• Reduce initial capital investment
• Distribute variable cost
expenditures over a drug’s
lifecycle

Gary.skarja@milliporesigma.com
Mochao.zhao@milliporesigma.com
Gary Skarja, Ph.D.
Mochao Zhao

Membrane Chromatography Solutions for Single-Use, Intensified mAb Purification

Recommended

Recommended

More Related Content

What's hot

What's hot (20)

Similar to Membrane Chromatography Solutions for Single-Use, Intensified mAb Purification

Similar to Membrane Chromatography Solutions for Single-Use, Intensified mAb Purification (20)

More from MilliporeSigma

More from MilliporeSigma (20)

Recently uploaded

Recently uploaded (20)

Membrane Chromatography Solutions for Single-Use, Intensified mAb Purification