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

10000

20000

30000

40000

50000

60000

70000

80000

0

5

10

15

20

25

30

35

40

45

50

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

22

: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

3

:14

:41

4

: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

11

:01

:22

11

:18

:02

11

:34

:42

11

:51

:22

12

:08

:02

12

:24

:42

12

:41

:22

12

:58

:02

13

:14

:42

13

:31

:22

13

:48

:02

14

:04

:42

14

:21

:22

14

:38

:02

14

:54

:42

15

:11

:22

15

:28

:02

15

:44

:42

16

:01

:22

16

:18

:02

16

:34

:42

16

:51

:22

17

:08

:02

17

:24

:42

17

:41

:22

17

:58

:01

18

:14

:41

18

:31

:21

18

:48

:01

19

:04

:41

19

:21

:21

19

:38

:01

19

:54

:41

20

:11

:21

20

:28

:01

20

:44

:41

Ine

rt C

ou

nts

Mic

rob

e E

qu

ival

en

t

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