Participate in the interactive webinar; http://bit.ly/SU-ADCWebinar Challenges of ADC manufacturing can be tackled by adopting single-use technology. Solutions will be presented about how to overcome concerns and implement a processing platform, supported through a strong vendor-manufacturer relationship. Explore our webinar library: www.emdmillipore.com/webinars

1. Merck KGaA
Darmstadt, Germany
Mary Robinette and Elizabeth Goodrich
September 26, 2019
Using Single-use
Technology to
Overcome the
Challenges of ADC
Processing

2. 2
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019
The life science business of
Merck KGaA, Darmstadt, Germany
operates as MilliporeSigma
in the U.S. and Canada.

3. Agenda
01 The Case for Using Single-use Equipment in
ADC Processes
02 Single-use Component Compatibility Testing
03 ADC Single-use cGMP Production - Lessons
Learned
04 Conclusions
3

4. Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019
The Case for
Single-use in ADC
Processes
01

5. 5
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019
Single-use Technology to Streamline Bioprocessing
Speed
Safety
Process
Economics
Flexibility
 Faster product changeover
 Accelerated time to clinic
or market
 Faster implementation
 Eliminates cleaning
Validation
 Reduced bioburden risks
 Reduced cross-
contamination risks
 Reduced operator and
environment exposure
 Adaptability to changing
process needs
 Multi-product
manufacturing facilities
 Scalable and movable
platforms
 Lower cost: reduced water/caustics,
utilities, equipment downtime, and
unproductive labor
 Compact footprint
 Lower investment

6. 6
Applicability of Single-use Technology for ADCs
 Process capability for ADC
processes
 Compatibility/Extractable Studies
including ADC solvents
 Single Use Assembly Testing
− Leak testing
− Class VI USP
 Product Specific Testing
− BioReliance® Extractable and
Leachable Validation Services
Solvent Component
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019

7. Single-use ADC Process
7 Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 20197
Reaction
Mobius® Mix
10-1000L
UF/DF
Mobius® FlexReady
TF2, TF3 2-10m2
Chromatography
Mobius® FlexReady
0.1 – 8LPM
Formulation/ Filtration
Mobius® Mix and Filters
10-1000L
Filtration
Opticap® Filter with Millipore Express®
SHC membrane
UF/DF
Mobius® FlexReady TF2,TF3
0.5-10m2
Solvent Component

8. 8 Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019
Solvent Compatibility Considerations
 Solvent composition
 Product contact components
 Performance testing
− Visual inspection for leaks and physical changes
− Functionality testing
− Sterile filters – verify microbial retention
− Extractables
ADC Single-use Process – Solvent Compatibility
T
Mobius® FlexReady Solution
with Smart Flexware®
Assemblies

9. Common ADC Process Solvents
 Drug charge to reactor:
− 100% DMSO (Dimethyl Sulfoxide)
− 100% DMAc (Dimethyl Acetamide)
 Reaction/Purification composition
− up to 20% DMSO
− up to 20% DMAc
ADC Single-use Process – Solvent Composition
Mobius® Single-use Jacketed
Mix 100 L
9 Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019

10. ADC Single-use Reactor – Solvent Compatibility
Mix bag components
 Bag film
 Ports
 Mixer
 Tubing
 Sampling valve
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201910
Mixer
Sampling
Mobius® MyWay
50L PureFlex™ Mobius® MIX Bag
Ports
Film
Example Mix Bag Components
PureFlex™ Film
Risk assess component exposure to 100% solvent

11. Flow path assembly components
 Pump
 Valves
 Instrumentation
 Mix bag
 Sampling
ADC Purification Equipment – Solvent Compatibility
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201911
Recovery Line
To Transfer Pump
Smart Flexware® Assembly for TFF
100L Single-use
Feed Container
Assembly
Filtrate Assembly with
Single-use Sensors Flow Cell
Retentate Sampling
Port Assembly
Dow Corning® Pharma-80 Tubing,
used to create bypass loop around TFF
liners and cassettes
Extractable Test Setup on Mobius® FlexReady Solution
with Smart Flexware® Assemblies for TFF
Single-use Feed
Pump Assembly
Test Setup on Mobius® FlexReady Solution with Smart
Flexware® Assemblies for TFF

12. Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019
Single-use
Component
Compatibility
Testing
02

13. After exposure to:
 20% and 100% Dimethyl Sulfoxide
(DMSO)
 20% and 100% Dimethyl Acetamide
(DMAc)
 Water
Provide understanding on compatibility of selected standard single-use assemblies and
components with the solvents most commonly used in ADC processing
Overview of Compatibility Testing
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201913
Evaluate functionality and extractables of:
 Mobius® MIX bag made from PureFlex™
film
 Opticap® XL filter with Millipore Express®
SHC membrane
 Mobius® FlexReady Solution with SMART
Flexware® assemblies for TFF or
Chromatography
 Lynx® CDR and Lynx® S2S sterile
connectors
 Colder AseptiQuik® G sterile connectors

14. PureFlex™ film
 100% DMSO, 100% DMAc, RO water
 24-hour soak in solvent at 40°C
Post-solvent exposure:
 No visual changes
 No change in bag seam strength
 Slight increase in film elongation at break
− No negative performance impact
expected
− In use, bag is supported in a holder
Mobius® MIX bag testing
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201914

15. Millipore Express® SHC 0.5 mm / 0.2 mm PES
membrane
 47 mm membrane discs and 5” Opticap® XLT
capsule
 20% DMSO, 20% DMAc, RO water
 24-hour soak in solvent at 40°C
Post-solvent exposure:
 No visual or physical changes
 Water flow passes specification
 Bubble point passes specification
 Capsule maintains complete microbial
retention (> 9 log reduction of B. diminuta)
Millipore Express® SHC membrane and Opticap® Filter Capsule testing
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201915
ADC Processing using Millipore Express® SHC
(Cat#KHGEG015FF3/Lot#C8DA57941)
Bacterial Retention Test
(B. diminuta;>107 CFU/cm2)

16. Opticap® Filter with Millipore Express® SHC Extractables Evaluation
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201916
 No pre-use flush of capsule
 All identified components are from membrane
manufacture or gamma irradiation
 In a worse-case process scenario with low
filter throughput (150 L/m2), all extractable
components would be < 0.1 mg/mL in pool
 Subsequent diafiltration will reduce even more
Opticap® XL5 Capsule with Millipore Express® SHC Membrane
Non-Volatile Extractables via HPLC-DAD/MS
Opticap® XL5 Capsule with Millipore Express® SHC Membrane
Semi-Volatile Extractables via DI-GC/MS Opticap® XL5 Capsule with Millipore Express® SHC Membrane
Volatile Extractables via HS-GC/MS

17. Flow Paths and Wetted Materials for Solvent Exposure Testing
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201917
Recovery Line
To Transfer Pump
Smart Flexware® Assembly for TFF
100L Single-use
Feed Container
Assembly
Filtrate Assembly with
Single-use Sensors Flow Cell
Retentate Sampling
Port Assembly
Dow Corning® Pharma-80 Tubing,
used to create bypass loop around TFF
liners and cassettes
Extractable Test Setup on Mobius® FlexReady Solution
with Smart Flexware® Assemblies for TFF
Single-use Feed
Pump Assembly
Lynx® Connectors Flow Loop AseptiQuik® G Flow Loop
Extractable Test Setup on Mobius ® FlexReady Solution
with Smart Flexware® Assemblies for TFF

18. Flowpaths for Post-Exposure Functional Testing
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201918
Valve Bypass Test setup #1
Valve Bypass Test setup #2
Valve Cycling Test setup

19. Metals Analysis
 Only traces of sodium and calcium were detected
 No metals identified are classified as elements to be considered in the risk assessment as outlined in
ICH Q3D Guideline for Elemental Impurities and USP <232> Elemental Impurities
Non-Volatile Organic Analysis
 No compounds were detected in any system or solvent combination using LC-MS analysis.
Volatile Organic Analysis
 HS-GC/MS analysis for volatile organic compounds indicated no extractable peaks in any system.
Semi-Volatile Organic Analysis
 DI-GC/MS analysis for semi-volatile organic compounds did not identify any peaks above 0.05 mg/mL.
 Results from DMAc exposure, shown on next slide, represent the highest observed levels of the three
solutions which were tested.
Mobius® Smart Flexware™ and Lynx® Connector Extractables
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201919

20. Extractables after Exposure to 20% DMAc
Smart Flexware® Assemblies and Connectors for Mobius®
systems, Method: DI-GC/MS
1 Mobius® TF-2S Smart Flexware® assembly, 6L recirculation
Extractables after Exposure to 20% DMAc: Direct Inject GC-MS
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201920
Extractables after Exposure to 20% DMAc
Smart Flexware® Assemblies and Connectors for Mobius®
systems, Method: DI-GC/MS
All identified compounds are < 0.05 mg/mL
1 TF-2S Smart Flexware® Assembly, 6L recirculation
1 Lynx® CDR and 1 Lynx® S2S, 0.5L recirculation
2 AseptiQuick® G connectors, 0.5L recirculation

21. Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019
ADC Single-use cGMP
Production –
Lessons Learned
03

22.  Single-use system design/suitability review:
end user and supplier
− User requirements collaboration session
− Proper selection of standard systems and
options
− Review options for single-use assembly
custom design
− Users attend supplier demonstration of single-
use equipment
− Outcome: optimal single use-system for
application
 Supplier training for production success
− Single-use system hardware
− Single-use assembly unpacking and installation
− Key to prevent single use assembly damage
ADC Single-use End-user / Supplier Collaboration for Success
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201922
Mobius® Single-use Jacketed Mixers
M Lab™ Burlington, MA

23. Single-use Technology Benefits - ADC Production experience
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201923
Speed
Safety
Process
Economics
Flexibility
 Faster product changeover
 Accelerated time to clinic
or market
 Faster implementation
 Eliminates cleaning
Validation
 Reduced bioburden risks
 Reduced cross-contamination
risks
 Reduced operator and
environment exposure
 Adaptability to changing
process needs
 Multi-product
manufacturing facilities
 Scalable and movable
platforms
 Lower cost: reduced
water/caustics, utilities, equipment
downtime, and unproductive labor
 Compact footprint
 Lower investment
Reduced ADC potent
waste handling/disposal

24. Platform ADC process equipment
 Standard automated hardware and single use
flowpath assemblies
− Examples: Chromatography and TFF skids, mix
bags
 Development of standards for custom designed
single use assemblies - multiple product/process
application
− Examples:
− Bag manifolds with pump tubing, sample bag
and filter in one single use assembly
− Mix bag assemblies for ADC reactor, process
blending and formulation
 Additional benefits
− Simplified inventory
− Reduce operator training/manual intervention
− Reduce autoclave parts
ADC Single-use - Production Experience
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201924

25. Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 2019
Conclusions
04

26.  Demonstrated solvent compatibility of the most commonly used ADC conjugation chemicals at
conditions that exceed typical concentrations and exposure times for:
− Mobius® Mix bag with PureFlex™ film
− Opticap® XL filter with Millipore Express® SHC membrane
− Mobius® FlexReady Solution with Smart Flexware® Assemblies for TFF or Chromatography
− Mobius Lynx® CDR, Mobius Lynx® S2S Connectors and Colder AseptiQuik® G sterile connectors
 After 24 Hour exposure to 20% DMSO or 20% DMAc
− No leaks and no physical changes observed by visual inspection
− Bags, flexware, connectors, and filters passed all functionality testing including microbial retention
− PureFlex™ film maintained performance after 24-hour exposure to 100% DMSO or 100% DMAc
 Extractables Profile
− No peaks were identified by LC-MS or HS GC-MS for any of the solvent extractions.
− Several peaks were visible using DI GC-MS. No compounds were present above 0.05 mg/mL.
− No metals identified by ICP that are classified as elements to be considered in the risk assessment
per ICH Q3D Guideline for Elemental Impurities and USP <232> Elemental Impurities
Conclusions: Compatibility and Extractables
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201926

27.  A broad range of single-use equipment is available to
meet the challenging needs of ADC production.
 Single-use assemblies are compatible with ADC process
solutions.
 Single-use supplier and end user collaboration is key
for successful process application.
 Single-use equipment benefits are realized in ADC
production.
 Use of standard and custom single-use systems allows
for an ADC process equipment platform that meets
multiple product/process needs.
Conclusions: Single-use Application for ADC Process
Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201927

28. Using Single-use Technology to Overcome the Challenges of ADC Processing | 26 September 201928
Acknowledgements
Compatibility and Extractables Testing Team:
Dana Kinzlmaier
Jessica Shea
Shannon Ryan
Venkata Raman
Alex Chalmers
Brian Callahan
Shikeba Shirzay
Mobius® Technical support
 R&D
 Applications Engineering
 Bioprocessing
Bio-Conjugation Center of Excellence St. Louis
 Global Process & Analytical Development
 ADC Production manufacturing

29. Director of MSAT Applications
Engineering
Email:
Elizabeth.Goodrich@milliporesigma.com
Principal Project Engineer, API Global
Engineering
Email:
Mary.Robinette@milliporesigma.com
Elizabeth GoodrichMary Robinette
The vibrant M, SAFC, Mobius, Lynx, Opticap, Millipore Express, Emprove, Flexware and PureFlex are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates. All other
trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources.
© 2019 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.
Thank you!