RAPID MICRO METHODS FOR MANUFACTURING
Jeffrey W. Weber, PAT Project Manager
Kalamazoo, MI
22 Jan 2014
2 IFPAC 2014
• Communications – MicroNet / Newsletter – Training
• Return on Investment • Site Support and Application Review • Technology Development / Consortium / Vendor
Collaboration – Development Cycle – Assessment, Development,
Transfer, Pilot, Routine Use – Peer-to-Peer Pharma consortiums and best practices – External presentations / Agency interactions
• Evaluation of New ARMMs – Vendor, literature review, and conference attendance
Pfizer RMM Team Introduction
3 IFPAC 2014
• Current Micro Testing
• Implementation guidance
• Perspective of new systems
• Successful RMM Platforms
– EndoSafe Experience
• Gaps and Opportunities
–Online Bioburden Water Analysis
Session Goals
4 IFPAC 2014
Traditional Micro Challenges
106
Microorganisms
103
Culturable
Microorganisms 1
Sampling
Limitations
Classic culture assay is less than perfect!
http://www.enviroliteracy.org/article.php/58.html
Risk Analysis
5 IFPAC 2014
Current Micro Lab Testing
Water 9%
API/Raw Mtls 1%
Finished 3%
Environmental Monitoring 85%
Other 2%
6 IFPAC 2014
• Discrete point sampling
– EM
– Raw Materials
– Personnel
– Sterility testing
• Limited temporal information
• Retrospective
– “all or nothing”
– Silos
Current Micro Testing
Product
People
Process / Plant
Raw Materials
Patient
7 IFPAC 2014
• Cycle Time Reduction
• Cost Avoidance
• Risk Reduction
• Increased Process Knowledge
• NOT Test Cost Reduction (based on current technology)
• Parametric Release
The quickest sterility test is the one you do not need to perform
RMM Drivers
8 IFPAC 2014
Patient
Raw Materials
• Continuous monitoring
• Holistic control
• Raw material characterization is part of process
• Extreme temporal information
• Infinite sublots
• Real Time Release Testing (parametric release)
Future Micro Testing
Product
People
Process / Plant
9 IFPAC 2014
Patient
Raw Materials
Future RMM Platforms
Product
People
Process / Plant
Automated plate reader
Aerosol cytometry
Liquid Bioburden
Automated Water Testing
Online Water BioBurden
Enhanced LIMS / SPC /
Cpk - MVDA
Personnel Automated Sampler
Enhanced COA data
10 IFPAC 2014
• Applications across several sites
• Compendial (required) testing
• High risk applications
• Examples
–Water
– Endotoxin
– Environmental Monitoring
– Sterility
Platform Roles
11 IFPAC 2014
Successful Platforms Technology: Chromogenic Limulus amoebocyte lysate, or
LAL assay. USP <85> and EP 2.6.14 Method D. A simplified and rapid endotoxin testing method.
Recommended Applications • Compendial endotoxin testing • Development studies • In-process testing System Roles • Water testing (routine and post-maintenance) • Buffer hold times • Cleaning verification • Endotoxin removal efficiency • Final Product testing
12 IFPAC 2014
• Online Total Organic Carbon for Water
–Minimal regulatory impact to online testing
– Improved process control over grab samples
• No guidance from USP, EP or JP initially for use of online testing
–ASTM guidance E2656-10
• Companies have moved to RTRt water release
–Business benefit
Model Implementations
13 IFPAC 2014
• RMM systems are orthogonal to compendial testing
• Different units
–CFU versus BioCounts
• How to react to new information?
–Regulatory requirement (i.e. heavy metals)
– Product Quality
• Interaction with Regulatory Agencies
RMM Challenges
14 IFPAC 2014
Manufacturing (PAT)
• In-plant testing
• Raw materials screening
• Process decisions
• EMA / EP support
QC (Micro Labs)
• Lean labs
• More automation
• Less “micro” training
• USP / FDA aligned
RMM Roles
RMM
15 IFPAC 2014
• Pfizer would like to see the development
–Online Water Testing
• At-line bioburden
–BioBurden Testing
• Simplified testing
–Online Endotoxin
– Environmental Monitoring
Industry Perspectives
16 IFPAC 2014
Regulatory Guidance
USP <1223> Validation of Alternative Microbiological Methods
EP 5.1.6 Alternative Methods for Control of Microbiological Quality
PDA TR No. 33 The Evaluation, Validation and Implementation of New Microbiological Testing Methods (update in process)
• Involve internal regulatory organization early in project
• Use pre-filing meeting and comparability protocols based on recent FDA presentations
17 IFPAC 2014
• Investigations
• Reduced in-process inventory
• Real-time testing and near-line, in-process use.
• Automation advantages lab resource reduction (training or technique).
–Human Error
• Special case applications are available.
RMM Benefits
18 IFPAC 2014
Pharmaceutical Waters
• 30 types defined
• 19 analytical waters
Purified Water
• TOC < 500 ppb
• Conductivity 1.4 μS @ 25 deg C
Water for Injection
• Bioburden < 10 CFU / 100 mL
• Endotoxin < 0.25 EU / mL
Pharmaceutical
Waters
Purified
Water
Water for
Injection
(food for bugs)
(bugs)
(bug parts)
Water Testing
19 IFPAC 2014
EndoSafe (Bacterial Endotoxin Testing)
20 IFPAC 2014
• Background & Compendia Test Options
• EndoSafe System Overview –pH
–Negative Controls
–PPC averages
– Sample Volume
• Quality Systems
• System Applications
• Summary
EndoSafe Overview
21 IFPAC 2014
Background & Compendia Test Options
22 IFPAC 2014
• Pyrogens are materials (natural or synthetic) that elicit a febrile (fever-inducing) response in the host.
• Bacterial endotoxin, lipopolysaccharides (LPS), are part of the bacterial wall of many gram negative bacilli.
– Bacterial endotoxins are normally released when the cell is lysed.
– Bacterial endotoxin produces the highest pyrogenic response.
Bacterial Endotoxin and Pyrogens
23 IFPAC 2014
USP <85> Bacterial Endotoxin Testing
• Gel-Clot
• Turbimetric
• Chromogenic
– Endpoint
–Kinetic
Compendia Testing
24 IFPAC 2014
EndoSafe System Overview
25 IFPAC 2014
EndoSafe Platform
• Two component System
• LAL impregnated cartridges
• EndoSafe Readers
• Portable Test System (PTS)
• Multi-Cartridge System (MCS)
26 IFPAC 2014
Cartridge
1 sample per cartridge, performs duplicate assays including positive product control
The sample passes through the channels, picks up and mixes with reagents allowing reading at the optical well of the equipment
Cartridges and reagents are stable for 1 year at 25 deg C.
27 IFPAC 2014
• Reduced operator steps
–Dilute and shoot
• Consistent calibrations
–Archived calibration curves
• Quantitative results
–Reduced laboratory investigations
Human Error Reduction
28 IFPAC 2014
• The EndoSafe system conforms to USP<85> monograph. – The standard curve is provided by vendor and
supported by the PPC. – The Positive Product Controls (PPC) provides
assay verification for each test – pH of the test specimen should be measured
to assure a pH within the range of 6.7 to 7.3. – Negative controls are monitored by facility's
incoming raw material testing program – Subject to product specific studies with quality
system controls; the PTS system conforms to USP<85>.
EndoSafe Conformance
29 IFPAC 2014
•PPC testing demonstrates the assay performs as designed and results should be distributed across the acceptance of 50% to 200% recovery. –Assay performance verification each time
Positive Product Control (PPC)
30 IFPAC 2014
•Periodic review of the PPC results by each product tested should not be grouped or centered at either end of the acceptable ranges. •Different dilutions of the sample solutions should not impact the PPC of the product.
PPC
31 IFPAC 2014
Positive Product Control
Average 100.5%
SD 20.67%
N=47,788
32 IFPAC 2014
Operator PPC
Average 99.6%
SD 20.25%
N=1,795
33 IFPAC 2014
Sample PPC
Average 99.2%
SD 17.46%
N=222
34 IFPAC 2014
Testing – Volume Study
• Pfizer performed confirmatory testing to assess sample volume impact – EndoSafe assay requires 25 μL
• Evaluated at 5, 10, 15, 20, 25, 30, and 35 μL with LRW – 5 & 10 μL no valid results
– 15 & 20 μL PPC samples results were high - too concentrated
– 30 & 35 μL had low PPC results, over diluted
• Vendor supplied 25 μL pipettor provides consistent results
2008 International Federation of Process Analytic Chemists (IFPAC)
Jeffrey W. Weber, Pfizer
35 IFPAC 2014
Testing – Volume Study
Target PPC value 0.76 EU/mL
0
100
200
300
400
500
10 10 15 15 20 20 25 25 30 30 35 35
Po
stiv
e P
rod
uct
Co
ntr
ol (
%)
Sample Volume (microliter)
36 IFPAC 2014
Quality Systems
37 IFPAC 2014
Quality Systems
• Standard Operating Procedures
• Consumable material control
• System Qualification
• Training
• Change Control
• Product Specific Validation
• Site Quality Oversight
38 IFPAC 2014
Three lots of material is evaluated:
pH of material 6.7 to 7.3
Sample CV < 25%
PPC CV < 25%
PPC recovery between 50% to 200%
– Inhibition evaluated at varied sample concentrations.
Product Specific Validation
39 IFPAC 2014
Pre-Checks – Check cartridge LOT#, Expiry Date & Sensitivity,
PTS/MCS calibration
• Printed results maintained in logbook which is reviewed by QA. – Technicians trained to enter results in LIMS.
– Key is that QC control the cartridges and release to Manufacturing
Post-Checks – Verify Time/Date on the printout is correct,
and matches LIMS Time/Date.
Pfizer Quality Systems
40 IFPAC 2014
Data Reporting
Raw Data
PPC Information
Sample Information
41 IFPAC 2014
System Applications in Pharmaceutical Manufacturing
42 IFPAC 2014
• Raw Material Testing
• WFI
–Point of use testing
–Post-maintenance release
• Buffers
• Cleaning Verification
• Final product release
System Applications
43 IFPAC 2014
• Point of Use
– Sample point flushed
– Sample collected
–Assay performed in plant
WFI Testing
44 IFPAC 2014
Online Bioburden Water Analyzer
45 IFPAC 2014
• Anthony Cundell (Merck)
• Oliver Gordon (Novartis)
• Nick Haycocks (Amgen)
• Joe Johnston (Fresenius)
• Michelle Luebke (Baxter)
• Neil Lewis (Procter & Gamble)
• Jeanne Mateffy (Amgen)
• Jeffrey W. Weber (Pfizer)
Online Water Bioburden Analyzer
46 IFPAC 2014
• Based on experiences of RMM and PAT systems; the workgroup is providing guidance to the industry and regulators.
• The workgroup has provided to instrumentation and water manufacturers;
–User specifications
– Testing protocols
–Business benefits estimates
OWBA Purpose
47 IFPAC 2014
Labor Reduction (Resource Allocations) – Decreased sampling frequency via on-line
monitoring and system optimization
Product Quality and Process Understanding – Reduced investigations to bioburden excursions
through continuous water monitoring – Increased process understanding and product safety
through real-time monitoring – Real time release of in-process intermediates,
process buffers/solutions and ingredient water
Energy Savings – Less frequent of sanitization cycles through
verification of system capability
OWBA Applications
48 IFPAC 2014
• Critical Control Points (CCP)
– System health and process understanding
• Point-Of-Use Testing (POU)
– Fit for use PRIOR to use testing
OWBA Implementation
Water Generation
TOC OWBA
Storage Tank
49 IFPAC 2014
0
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50000
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80000
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8:3
8:0
7
10
:11
:27
1
1:4
4:4
7
13
:18
:07
1
4:5
1:2
7
16
:24
:46
1
7:5
8:0
6
19
:31
:26
2
1:0
1:2
6
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:34
:46
0
:08
:06
1
:41
:25
3
:14
:45
4
:48
:05
6
:21
:25
7
:54
:45
9
:24
:45
1
0:5
8:0
5
12
:31
:24
1
4:0
4:4
4
15
:38
:04
1
7:1
1:2
4
18
:44
:44
2
0:1
8:0
4
21
:48
:04
2
3:2
1:2
3
0:5
4:4
3
2:2
8:0
3
4:0
1:2
3
5:3
4:4
3
7:0
8:0
3
8:4
1:2
2
10
:11
:22
1
1:4
4:4
2
13
:18
:02
1
4:5
1:2
2
16
:24
:42
1
7:5
8:0
1
19
:31
:21
2
1:0
1:2
1
22
:34
:41
0
:08
:01
1
:41
:21
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:14
:41
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:48
:00
6
:21
:20
7
:54
:40
Ine
rt C
ou
nts
Mic
rob
e E
qu
ival
en
t
Bio Cell Cnt Inert Cnt
Heating to 65 deg C
Three day evaluation
Compendial Limit 10 CFU/100 mL
Likely microbubbles caused by
pressure drop from line of 30 psig
to 4 psig at the unit –
demonstrated by the decrease
during water heating cycles.
Data from Instant BioScan RMS-UM
50 IFPAC 2014
0
5000
10000
15000
20000
25000
30000
35000
40000
45000
50000
0
2
4
6
8
10
12
8:4
8:0
2
9:0
4:4
2
9:2
1:2
2
9:3
8:0
2
9:5
4:4
2
10
:11
:22
10
:28
:02
10
:44
:42
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:01
:22
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:18
:02
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:34
:42
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:51
:22
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:08
:02
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:24
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:41
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:58
:02
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:14
:42
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:22
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:48
:02
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:04
:42
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:21
:22
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:38
:02
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:54
:42
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:11
:22
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:28
:02
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:44
:42
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:01
:22
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:18
:02
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:34
:42
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:51
:22
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:08
:02
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:24
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:58
:01
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:14
:41
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:31
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:48
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:04
:41
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:21
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:38
:01
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:54
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:11
:21
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:28
:01
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:44
:41
Ine
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Mic
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19-May-13
Bio Cell Cnt Inert Cnt
Routine Operation
Heating to 65 deg C
Data from Instant BioScan RMS-UM
51 IFPAC 2014
Assay Daily Weekly Two Week
Interval
Single Test
Point
Bioburden
($65 to
$110/sample)
$24,000 to
$40,000
$3300 to
$5700
$1600 to
$2860
Estimated annual cost for bioburden sampling, testing and data reporting
52 IFPAC 2014
• “No savings for any company until acceptance and implementation”
• The workgroup members will support external collaborations with other manufactures and vendors with the goal of presenting data to a wider forum.
• Longer term roles for the system may include; – pre-sterilization in-process bioburden – water system qualification – filling line monitoring – investigations.
OBWA Next Steps
53 IFPAC 2014
• RMMs are being implemented across the manufacturing spectrum
–Business benefits
–Mature systems are being available
• Manufacturing base is providing vendor feedback and collaboration
• Most of the roadblocks are “red-herrings”
–Replication and standardized systems.
Conclusions
54 IFPAC 2014
• Steve Hammond
• John Ruesch
• Dennis Jones
• Kevin Ryan
• Mike Baumstein
• Hilary Chan
• Thuy Bui
• Amy McDaniel
• Heather Mingia
• Joanny Salvas
• Aisling Lawton
• Cristian Sammartino
• Gabriella Caldarera
• Brandye Michaels
• Jerry Ryan
• Susan Berlam
• Michael Fenster
Acknowledgements
55 IFPAC 2014
Questions?
56 IFPAC 2014
TOC 500 ppb carbon
1.4 μS @ 25 deg C
BioBurden
10 cfu / 100 mL
Endotoxin
0.25 EU / mL
500 x 10-9 g /L
(nanograms)
5 x 10-12 g /L
(picograms) 4.3 x 10-13 g / cell
E. Dalahaye et al.
Water Research (2003)
1 x 10-15 g /L
(femtograms)
[Carbon] in Waters