Implementing and Managing Pre-use Post-sterilization Integrity Testing (PUPSIT)

This presentation explores best practices and case studies in aseptic processing, including how to implement and manage PUPSIT. You will learn:

• Integrity Testing – the background on IT itself, why it is important, and how it works
• Filtration setups and single-use technology
• The PUPSIT debate and how PUPSIT can be achieved with current technology, final filling, formulation, filtration

To learn more about this topic or collaborate with our technical experts, schedule a remote visit at our M Lab™ Collaboration Centers: www.merckmillipore.com/remotevisit

1. Merck KGaA
Darmstadt, Germany
Dr. Paul Beckett
Technology Manager, EMEA
Guillaume Lesage
Technology Consultant, EMEA
Joint Pharmaceutical Analysis Group (JPAG) Aseptic
Processing Conference, May 9, 2019
Practical Examples of Overcoming Common Challenges
Implementing and
Managing PUPSIT

2. 01
02
03
Agenda
Case studies
•What went right?
•What went wrong?
2
Integrity Testing
•The background on the IT test itself, why it is
important, and how it works
Filtration Setups and Single-Use Technology
•Options when it comes to designing filtration
trains for final fill operations into isolators or
RABS.
PUPSIT
•The PUPSIT debate and how PUPSIT can be
achieved with current technology

3. Check correct installation
•Detects system leaks due to o-rings, gaskets, faulty
seals
Confirms manufacturer’s specifications
•Assures the correct pore size filter
Check for damages
•Assures integrity before/after sterilization
Sterility assurance
•Sterile filter is at the core of the aseptic process
Regulatory requirements
•Link between validation and current processing
conditions
•GMP Requirement
Why Perform Integrity Testing?
3

4. Defect
made visible
by smoke
Virus retention
Bacteria retention
Aerosol
Air binary gas
Freon Diffusion
Current flow test for PTFE
Air Diffusion
Bubble point, WIT
▪Water
▪Low surface tension fluid
Integrity Test Sensitivity
Sensitivity
Proprietary
High sensitivity
Industry
Standards
In development
Gold standards
4

5. Physical Integrity Test
Diffusion + Bubble point
Gas
Flow
Bubble
Point
Pressure
Diffusion
Viscous flow
Viscous flow
Bubble point
Pressure
5
Use the right test!

6. Correlation to bacteria retentionQuantifying Retention Performance
Graphical SummaryBubble Pt.
O
b
s
e
r
v
e
d

C
F
U
6050403020
3000
2500
2000
1500
1000
500
0
Retentive?
No
Yes
Measure Retention
Versus Bubble Point Bubble Pt.
L
o
g
(
C
F
U
)
6050403020
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
Use the Relationship
Log(CFU) vs. Bubble Pt. Bubble Pt.
R
e
t
e
n
t
i
o
n

C
o
n
f
i
d
e
n
c
e

(
%
)
6050403020
<1
37
90
99
99.9
99.99
Determine Bubble Point
with High Retention Confidence
Bubble point can have
a direct correlation
Diffusion & other tests can
have an "go -no go"
correlation
# of
Samples
Retentive
Non-
Retentive
246810 12 14 16 18 20 22
24
2
6
Specification
Diffusion rate (cc/min)
6

7. Supplier
End
-
User
Integrity Testing
A critical element of overall sterility assurance strategy
Validated sterilizing membrane
Validated sterilizing filter
Bacterial retention validation
testing (WCC)
Process Validation and Control
Batch integrity test
7

8. 01
02
03
Agenda
Case studies
•What went right?
•What went wrong?
8
Integrity Testing
•The background on the IT test itself, why it is
important, and how it works
Filtration Setups and Single-Use Technology
•Options when it comes to designing filtration
trains for final fill operations into isolators or
RABS.
PUPSIT
•The PUPSIT debate and how PUPSIT can be
achieved with current technology

9. Filtration
design options

10. Regulatory Compliance
Reference: EudralexVolume 4 Annex I

11. Formulation, Filtration and Final Filling
ProcessOverview
Flush Bag
or
Barrier
Filter
Vent Bag
Filter Capsule
(Single/Redundant)
Filtration
Sterile connector
Sterile disconnect
Bulk Fluid Bag
Or Formulation
Vessel
Pump
Sampling
Formulation
Header Bag
(Surge)
Filling Needle(s)
Pump
Filling
All components can be
designed in SUT
Hybrid solutions are
widely used (single-use
and multi-use
components)
Sterileconnector is a key
component to connect
the process steps
Finalfilling step under
cleanroom grade A
conditions

12. There are a variety of final filter set-ups in use
Final Sterile Filtration Options
Dual filter
Bioburden /
Sterile Filtration
Sterile –Hold -
Sterile Filtration
Redundant and
Series
Sterile Filtration
•Compliance with regulatory guidance for <10 cfu/100 ml
•Very low plugging risk for primary filter
•Potential batch recovery if one filter fails IT
•Higher hold up
•Higher cost
•Highest system complexity
•Compliance with regulatory guidance for <10 cfu/100 ml
•Very low plugging risk for primary filter
•No back-up in the event of primary filter failure
•Higher hold up
•Higher cost
•Higher system complexity
Single filter
Sterile Filtration
•Minimum hold-up volume
•Minimum flushing requirements
•Ease of handling and operation
•Lower filter cost
•No back-up in the event of primary filter failure
•Feed bioburden control
•Filter plugging

13. Single Filter
•Low value products
•Very low volume products
•Product can be easily reworked
•Processing of bulk drug substance

14. Dual Filter
•Products coming from open processing
conditions
•10 CFU/100mL is a requirement!
•Bioburden reduction filter is also sensible
risk management
•Only one of the filters needs formal
bacterial challenge validation

15. Redundant Filtration
•Usually has a prefilter too
•Both filters are validated by bacterial
challenge
•Only ONE filter needs to pass testing for
the batch to be released
•Business risk management
•Very common for reasonably sized fills of
high value products

16. Serial Filtration
•The same as redundant filtration in layout
•However, both filters are validated together
•BOTH filters need to pass testing to release
the batch
•Commonly used in applications where
formulation components weaken correlation
between bubble point and bacterial retention

17. What connects
the filters to
the filling line?

18. Increase filling productivity –reduce time to market
Why Single-Use Technology in Aseptic Processing?
Reduced risk of cross contamination
Avoids aseptic assembly
Validated sterilization process
Minimize capital investment (no
CIP/SIP)
Outsource sterilization & assembly
Reduce laborand on-going
validation burden
Achieves high flexibility (multi
purpose line)
Facilitates contract manufacturing
Fast small scale clinical
manufacturing
Process Security Enhanced Economics Speed-to-market
1 2 3
Mobius
®
single-use assembly installed in GroningerFlexPro

19. Ability to interface with an isolator while maintaining sterility
Isolator Interface with GetingeDPTE
®
Beta Bag
Sterile transfer of fill lines and needles

20. Easier handling of filter placed in grade C
Sterile Filter Placement
Grade C
Grade A
(RABS/
isolator)
Bulk
Product
Peristaltic
Pump
Sterile
Filter
Header
Bag
Peristaltic
Pump
Filling
Needle
Grade C Grade A (RABS/isolator)
Bulk
Product
Peristaltic
Pump
Sterile
Filter
Header
Bag
Peristaltic
Pump
Filling
Needle
Filter in Grade A /
ISO 5 / Class 100
Low risk to fluid path
contamination
Open vents, connections
Sterile environment
risk from vented
liquid
Handling challenges
Large number of
components to transfer
Pre-use integrity testing
challenge
Filter in Grade C / ISO 7
/ Class 10,000
Easier manipulations
Installation, flushing,
integrity testing, venting,
pump tubing adjustment
Potential to replace filter
if integrity fails
More potential points for
ingress
Demonstration of
assembly integrity more
critical

21. 01
02
03
Agenda
Case studies
•What went right?
•What went wrong?
21
Integrity Testing
•The background on the IT test itself, why it is
important, and how it works
Filtration Setups and Single-Use Technology
•Options when it comes to designing filtration
trains for final fill operations into isolators or
RABS.
PUPSIT
•The PUPSIT debate and how PUPSIT can be
achieved with current technology

22. 22
The Debate-PUPSIT

23. 23
The Guidance (As It Stands)

24. PUPSIT in the most complex assembly
24
Primary Product Filter
Product Inlet
Redundant Product Filter
SPG GasFilter
Barrier Filter
Flush/AirInlet
VentCollection Bags
Product Outlet

25. 25 25
PUPSIT is more than possible with single -use assemblies

26. Millipak
®
and Millidisk
®
Barrier Filters
Liquid
Wetting and
flushing fluids
evacuate to
drain
Gas
Air from
integrity test
and blow
down can
vent from
same filter
One disk pair
3 Hydrophilic
membranes
1 Hydrophobic
membrane

27. Redundant Filtration –Enable a Valid Test
Pre-use
F1 F2
P2
P1
Post-use
F1 F2
P2
P1
optional
27

28. Case studies

29. Get the Filter Sizing Right
Small volume fill
10L BDS of a biologic
Processed through a 10” PVDF 0.22um
No flushing –E&L
Hold up concerns
BDS
10L
Durapore
®
membrane
0.22um
10”
Isolator boundary
IT
Durapore
®
membrane
0.45um
10”

30. Tricky products
Viral vectors are booming
25L of lentiviral vector
60% yield loss
Closed system design
0.45um/0.45um
BDS
25L
Durapore
®
membrane
0.22um
Millipak
®
filter
200
Isolator boundary
IT
Durapore
®
membrane
0.45um/0.22um
Millipak
®
filter
200

31. Policy is Inside the Isolator
Checking the dimensions
Actually from CCM filtration
Expensive hardware for storage
Capsule would not fit through the port
Same concerns with too much tubing –
integrity tester inside the isolator
BDS
Millipore
®
Express SHR
XL300
0.1um
Isolator boundaryIT
Millipore
®
Express SHC
XL300
0.5/0.2um

32. Devil is the Detail
Difference between aseptic and sterile
Assemblies can be very big, too big to
ship
They need to be connected, sometimes
this connection needs to take place in the
isolator
Ensure this connector is up to standard
Transfer connector to outside the isolator
BDS
Durapore®
membrane
0.22um
Millipak® filter
200
Isolator boundary
IT
Durapore®
membrane
0.45um/0.22um
Millipak® filter 200
Aseptic connector

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