1.overview Of Bioprocess And Regulatory Requirements For Filtration Of Biological Products.pdf

OVERVIEW OF BIOPROCESS AND REGULATORY REQUIREMENTS FOR FILTRATION OF BIOLOGICAL PRODUCTS Microbiological Monitoring, Risk Assessment, Microbiological Control and Filter Qualification Overview

Michael Payne Merck Millipore

Impact of microbiological contamination Routes of contamination in the process Risk assessment and mitigation strategies Filter categorization Moderately critical filters and risk approach Critical filters and risk approach

Filter qualification

Overview of Generic Biological Manufacturing Process MCB

WCB

Seed Train Buffer

Buffer

Raw Material s

Buffers

Harvest

Buffer Buffer

Buffer Buffer

In-Process Contamination

Biologics

30 Percent Average percent of process deviations caused by contamination*

1- 6 Months

Length of time to complete an investigation

1-10

Million Euro

Average operations cost of microbial contamination

Impact: Interruption of patient product supply, delays in clinical development, batch loss, consent decrees, requalification studies, financial losses

4

*Sources Langer 2013, Wiebe 2014

Impact of microbiological contamination Routes of contamination in the process Risk assessment and mitigation strategies Filter categorization Moderately critical filters and risk approach Critical filters and risk approach

Filter qualification

3D System Risk Assessment Concept

Used to calculate practical severity

Considers

• System‘s distance from the process stream • Location along the process stream • System‘s complexity

Highest score is highest risk

(proximity x location x complexity)

This tool is mainly used to assign a risk level to an overall system before assessing failure frequency and detectability (SOD / RPN) Excellent for complex systems as part of “big picture” analysis to prioritize risk management “A 3-D Risk Assessment Model”, Journal of Validation Technology [Autumn 2008] pp70 - 76

Overview of Generic Biological Manufacturing Process Microbiological Risk

MCB

WCB

Seed Train Buffer

FOCUS

Buffer

Raw Material s

Buffers

Harvest

Buffer Buffer

Buffer

Buffer

Regulatory and Compliance Microbiological FOCUS

MICROBIOLOGICAL RISK HAMMOCK

Contaminants of Concern in Biologics

Adventitious Agents Microorganisms that have been unintentionally introduced into the manufacturing process of a biological product:

 Bacteria  Fungi  TSE Agents  Virus

HIV infecting a human T cell © NIBSC/Science Photo Library

Routes of Contamination MCB

WCB

Seed Train Buffer

Buffer

Raw Material s

Buffers

Harvest

Buffer Buffer

Buffer Buffer

Routes of Contamination

Seed Train Contamination with Mycoplasma MCB

WCB

Seed Train Buffer

Buffer

Raw Material s

Buffers

Harvest

Buffer Buffer

Buffer Buffer

Routes of Contamination

Bulk solution contamination with Bacillus MCB

WCB

Seed Train Buffer

Buffer

Raw Material s

Buffers

Harvest

Buffer Buffer

Buffer Buffer

Key Points

Routes of Contamination Many routes for microbial contamination  Increased awareness of virus, mycoplasma, and Leptospira contamination in upstream processes

Intensive risk assessments could have prevented many of these contaminations Raw Materials are a significant cause of contamination Downstream contamination is often the result of:  Improper cleaning or sanitization  Suboptimal system design

Leverage supplier expertise during process development

Impact of microbiological contamination Routes of contamination in the process Risk assessment and mitigation strategies Filter categorization Moderately critical filters and risk approach Critical filters and risk approach

Filter qualification

Risk Assessment to Prevent Contamination

Identify

Mitigate

Detect

Risk Assessment: Identify

Identify

Mitigate

Detect

Assess extent of risk, ability to detect, and frequency of occurrence

Identify Minute virus of mice (MVM) ~18-24 nm

Facility Equipment Process Materials

Acholeplasma laidlawii < 0.2 µm

Each source is a potential entry point for microbial contamination Leptospira species 0.4 µm x >>5 µm

Utilities Personnel Bacillus species 1 µm x 4 µm

Case Studies of Microbial Contamination in Biologic Product Manufacturing Suvarna, K., Lolas, A., Hughes, P., Friedman, R. Biotechnology Manufacturing Team, Division of Manufacturing and Product Quality, Office of Compliance, Center for Drug Evaluation and Research, Food and Drug Administration

Process Flow Raw Materials

Each step may introduce microbes into the process Handling

Water transfer (cleaning, compounding)

Transport of materials in the facility

Compounding

Testing

Mixing

Sampling

Hold times

Transfer into different packaging

Dispensing

Storage conditions

Sampling

Weighing

Room Cleaning

Sieving

Equipment Cleaning

Crushing

Personnel Hygiene

Sifting

How do I assess the risk of these parameters?

What process inputs could introduce contamination

Example: Cause and Effect Diagram – the 6 “M”s (Hu)Man

Material

Mother Nature Air Handling

Hygiene

Transfer

Training Aseptic Technique

Water Tubing / Piping Mineral Salts Filters

Insects Animals

Humidity

Single use Microorganisms Basal Medium Supplements Tanks Packaging Pallets Reagent

Sieves Crusher

Pump Autoclave

Water System Load cells

Steam Generator Mixer

Drum Lifter Pump

Air compressor

Machine

Mixing Compounding

Transfer Testing

Hold Time Sampling Room Cleaning Weighing Dispensing Filtration

Method

Calibration Microscopy Pressure Time

Flow Rate Counting

Temperature Quality Parameters

Measurement

Risk Assessment: Mitigate

Identify

Eliminate source or reduce likelihood of occurrence

Mitigate

Detect

Prevent Human Contamination

Strategies for prevention, mitigation and detection Prevention  Remove people from the environment

Mitigation When people have to be in the environment  Wear cleanroom attire  Work in cleanrooms

 Properly trained personnel

Detection  Viable air sampling

 Surface monitoring  Personnel monitoring

Prevent Raw Material Contamination

Raw Material Selection

Prevent  Remove animal derived components  Caution! Serum-free does not mean mycoplasma free  Consider chemical free  Recombinant alternatives to serum  r-Insulin, r-Transferrin & r-albumin  Select raw material quality grade  Pharmaceutical grade versus analytical grade  Audit vendor

Mitigate  Pre-treat components  Choose treatments effective for viral and bacterial reduction

Detect  Screen raw material with rapid tests – Caution! Sample sizes versus kG to tons of material

Mitigate Contamination

Raw Material Pre-Treatment Robust Clearance

Media Compatibility

Point of Use

Scalability

HTST

Yes

Component dependent

Yes

Large Scale

UV-C

Microorganism dependent

Component dependent

Yes

Challenging at large scale

Yes but challenges at large scale

Component dependent

No

Small batches

Yes

Yes

Yes

Yes

Yes

Yes

Not downstream small virus filters. Yes upstream virus filters

Technology

(~102C ~10 sec)

(254 nm)

Radiation

Mycoplasma Filtration

Virus Filtration

Yes If specifically claimed consistent LRV Yes by size exclusion consistent LRV

Yes

Yes

Cost Effective Yes at Large Scale

Key Points

Mitigate Prevention  Best option wherever possible

Containment  Personnel Control  Single Use Technologies

Raw Material Selection  Vendor qualification

 Pre-treatment

Downstream Processing  Viral Clearance  Filtration  Sanitization, cleaning and storage

Risk Assessment: Detect

Identify

Mitigate

Detect

Determine location, frequency, and limits of detection

Sterile or virus-free is only as good as the detection method used

Microbiological Detection Classical Methods

Rapid Methods

Most developed in the 19th century

Developed over the past 30 years but slow adoption rate

 Microscopy  Growth-based methods Benefits  Easy to implement  Easy to qualify

 Larger sample volumes possible Limitations  No universal medium or growth conditions  Only detect those microbes capable of replicating in the chosen test medium under the specified conditions  Can take days to weeks for a result

 qPCR  TMA  Microcolony growth detection Benefits  Rapid results  Higher sensitivity for equal volume compared to classical methods Limitations  More extensive validation  Higher expertise required  False positives doesn’t distinguish viable cells  Small sample size  Often destructive  Split samples needed for identification

Limits of Detection

Sampling Volumes

Sampling  Vessel Liters to 10,000+ Liters  Sample Volume  Less than 1 Liter

Assay  Removed from sample volume  Milliliter to microliter

Limits of Detection

Sampling Assume a 1 L sample from a 10,000 L Bioreactor Assay requires a 1 mL sample for testing CFU per Liter

10

1,000

10,000

CFU per mL

0.01

1

100

Probability an organism will NOT be detected in the sample

0.99

0.9

0.37

Assay Sensitivity LOD PCR for Leptospira:

100 CFU

LOD PCR for Mycoplasma:

1-10 CFU (equivalent)

LOD by light microscopy @ 400 x:

105 to 106 cells

LOD TCID50:

15 to 104 TCID50/mL

(equivalent)

Risk Assessment to Prevent Contamination

Identify

Eliminate source or reduce likelihood of occurrence

Assess extent of risk, ability to detect , and frequency of occurrence

Mitigate

Detect

Determine location frequency, and limit of detection

Impact of microbiological contamination Routes of contamination in the process Risk Assessment and Mitigation Strategies Filter locations and microbiological concerns Moderately critical filters and risk approach Critical filters and risk approach

Filter qualification

3D System Risk Assessment Concept Considers

a system‘s distance from the process stream its location along the process stream the system‘s complexity

Highest score is highest risk This tool is mainly used to assign a risk level to an overall system Excellent for complex systems as part of “big picture” analysis

“A 3-D Risk Assessment Model”, Journal of Validation Technology [Autumn 2008] pp70 - 76

Overview of Generic Biological Manufacturing Process MCB

WCB

Seed Train Buffer

Buffer

Raw Material s

Buffers

Harvest

Buffer Buffer

Buffer

Buffer

Regulatory and Compliance Microbiological FOCUS

MICROBIOLOGICAL RISK HAMMOCK

For illustrative purposes. Regulatory requirements, industry guidance and previous data will factor into a microbiological sampling plan.

Biopharmaceutical Process

Upstream

Sterility (Claim)

Bioburden Virus Mycoplasma

Working Cell bank

Seed Train

Endotoxin

Raw Materials Water Media Supplements pH Adjusters Antifoam

Media Prep

Mixing Pre-treatment Filtration

Production Reactor > 500 L

Clarification

Filtration Centrifugation

Harvest

Control Strategy for Upstream Processes

From A-Mab 33

Biopharmaceutical Process

Downstream Purification (1)

For illustrative purposes. Regulatory requirements, industry guidance and previous data will factor into a microbiological sampling plan.

Sterility (Claim) Bioburden Virus Mycoplasma Endotoxin

Buffer Preparation Filtration

Product Capture Filtration Harvest

Affinity Chromatography (Protein A)

Concentration Process Limit < X CFU (~ 20/ml)

Filtration

Intermediate Bulk Process Limit < 10 CFU/ml

Biopharmaceutical Process

Downstream Purification (2)

For illustrative purposes. Regulatory requirements, industry guidance and previous data will factor into a microbiological sampling plan.

Buffer Preparation Filtration

Intermediate bulk Process Limit < 10 CFU/ml

Filtration

Virus Inactivation Detergents Solvent pH

Sterility (Claim)

Chromatography Cation exchange

Filtration

Process Limit < X CFU (~ 20/ml)

Bioburden Virus Mycoplasma

Concentration

Small Virus Filtration

Biopharmaceutical Process

Downstream Purification (3)

For illustrative purposes. Regulatory requirements, industry guidance and previous data will factor into a microbiological sampling plan.

Sterility (Claim) Bioburden Virus Mycoplasma Endotoxin

Filtration

Chromatography Filtration

Concentration

Filtration

Bulk

Anion exchange

Process Limit < X CFU (~ 20/ml)

Bulk Drug Substance In Single Use Containers

Specification < 10 CFU/ml

Control Strategy for Downstream Processes

From A-Mab 37

Filters in the Formulation / Filling Suite Vent filter

Vent filter

WFI

Vent filter

IT gas inlet filter

Blanket / Transfer Gas Filter

Washing filters

Vial Washing

CIP

Vent filter

IT gas inlet filter

Protection filter

API

Drying filter

Clean Room Utility Gas Filters Protection filter

Depyrogenation

Gassing Vent filter Filter

WFI

Excipient

Formulation

38

Sterile Filtration

Prefilter Bioburden Sterilizing Filter Filter Bioburden & Sterile Filtration

Sterile Hold Tank

Stopper Washer Dryer

Aseptic Filler

Autoclave

Freeze dryer

Control Strategy for Drug Product Processes

From A-Mab 39

Impact of microbiological contamination Routes of contamination in the process Risk assessment and mitigation strategies Filter categorization Moderately critical filters and risk approach Critical filters and risk approach

Filter qualification

Filter Definitions Service

 The filter does not affect product quality  Where process fluids come from facility-wide systems, are not tailored to a specific process and do not have contact with the drug substance or potential drug substance.  Part of a No-Impact System - Where the equipment of system has no impact, direct or indirect, on product quality (ISPE Commissioning & Qualification Baseline Guide (2001))  Examples: distribution gas filter, water prefilter

Moderately critical  The filter indirectly affects product quality  Where process fluids “will not be in direct contact with exposed sterile product or surfaces.” (PDA TR40)  Part of an Indirect Impact System - equipment or system expected to have incidental or secondary impact on product quality (ISPE Commissioning & Qualification Baseline Guide (2001))  Examples: vent filter in a grade D/C area, bioburden reduction filter

Critical Applications  The filter directly affects product quality  Where process fluids “are in direct contact with sterile final product or critical surfaces of the associated equipment.” (PDA TR40)  Part of Direct Impact System - equipment or system that will have focused and immediate impact on product quality (ISPE Commissioning & Qualification Baseline Guide (2001))  Examples: vent filter on a sterile hold vessel, sterile liquid filter

Advantages of Classification - Multiple Process Lines or Bioreactor Trains

Categorize each filter in a line based on risk, then duplicate across the whole production area

Understand impact of language – Critical? Challenge can be to identify and agree on critical applications Example questions

– Does the fluid come into contact with a sterile surface? - Where is the filter position in relation to downstream purification - In what room classification is the filter prepared and used?

- How is the filter prepared for use? - What is downstream of this process step? - What quality testing is done on the filtrate?

Understand impact of language – Moderately Critical?

Challenge can be to identify and agree on moderately critical applications – and then on the degree of “moderate” Example questions – Where is the filter position in relation to downstream purification? - In what room classification is the filter prepared and used? - How is the filter prepared for use? - What is downstream of this process step?

- What quality testing is done on the filtrate? - What happens when the filtrate fails microbiological testing?

Impact of microbiological contamination Routes of contamination in the process Risk Assessment and Mitigation Strategies Filter categorization Moderately critical filters and risk approach Critical filters and risk approach

Filter qualification

Purpose of Moderately Critical Filtration

Removal of undesirable microorganisms from process fluids  Prevent contamination of the fermentation  Cell culture media and air  Formulation and process tanks  Chromatography systems  Buffers, washing fluids  Process intermediates

Reduction of bioburden in purification process steps  Low bioburden means low endotoxin  Low / controlled / specified bioburden may be a compliance requirement

46

Buffers in a Generic Biologicals Process - moderately critical filtration

Closer to the formulation point, the higher the risk

47

Risk

Example of a Moderately Critical Filter: Buffer filtration for the chromatography step:

Particulates

Microrganisms

More column cleaning

48

Block the flow distributor

Contaminate the chrom media

Longer run time

Change the column operation efficiency

Product variation / deviation issue

Reduce the life cycle of the media

Increased Cost

Exceed the validated cycle time

Deviation & Quality Issue

Some Filter Risk Assessment Considerations Contact time

Fluid classification

The longer the contact time the greater the risk

Fluids labeled “sterile” have the highest risk

Process conditions

Dosage form

The more aggressive the conditions, the greater the risk

Injectables without preservative have highest risk

Room classification Lower grade brings greater risk if there is a breech

Location of filter in the process The closer to the final product the greater risk

Detectability of poor filtration performance No in-line testing has the highest risk

Fluid pretreatment Less pretreatment has greater risk

Fluid posttreatment No downstream removal of low MW material has greater risk

Filter pretreatment

The more aggressive the pretreatment (e.g. SIP), the greater the risk

Prior history If there have been previous filter related issues, the risk is greater

Impact of microbiological contamination Routes of contamination in the process Risk assessment and mitigation strategies Filter categorization Moderately critical filters and risk approach Critical filters and risk approach

Filter qualification

Retention: What are the requirements for sterile medicinal products “All Sterilization Processes Should be Validated.”

WHO Annex 6: Good Manufacturing Practices for Sterile Pharmaceutical Products

Integrity Testing

Binding

Integrity Testing

Fit for Use Duty

Retention

QS, VMP & Retention

Stability

Documentation Sterilization

section 5.4 page 273

“Whatever type of filter or combination of filters is used, validation should include microbiological challenges to simulate “worst case” production conditions. The selections of the microorganisms to perform the challenge test (e.g. P. diminuta) has to be justified. The nature of the product may affect the filter and so the validation should be performed in the presence of the product……”

PIC/S Guide for Inspectorates: Recommendation on the Validation of Aseptic Processes

Compatibility Extractables & Leachables

A summary should be provided containing information and data concerning the validation of the retention of microbes and compatibility of the filter used for the specific product. US FDA Guidance on Sterilization Validation

51

What Critical Filters need to be Qualified for a Sterile Medicinal Product

Sterilizing liquid filter

Bioburden reduction filter Sterilizing gas filtration But not all filters need to be qualified in the same way or in the same depth

Generic Sterile Formulation / Filling Suite - Old style sterile filtration system with BBR

and EMA compliant

Vent Filter

Vent Filter

Prefilter

Formulation

Bioburden Reduction Sterilizing Filter Filter

Sterilizing Filter

Sterile Hold Tank

Aseptic Filler

What filters need to be pre-use integrity tested? What filters need to be post-use integrity tested? What happens if the “final filter” fails post-use integrity testing?

Generic Sterile Formulation / Filling Suite - Old style sterile filtration system with BBR.

EMA compliant, and FDA compliant for at risk product

Vent Filter

Vent Filter

Prefilter

Formulation

Bioburden Reduction Sterilizing Filter Filter

Sterilizing Filter

Sterile Hold Tank

Sterilizing Filter

Aseptic Filler

What filters need to be pre-use integrity tested? What filters need to be post-use integrity tested? What happens if the “final filter” fails post-use integrity testing?

How can I put the risks into perspective?

Example: Failure Mode and Effects Analysis (FMEA)

Score the parameters    

Probability of occurrence Severity Ability to detect Criticality

Prioritize the parameters

CMC Biotech Working Group: A-Mab: A case Study in Bioprocess Development http://www.casss.org/?page=286

Example of 3D Risk Assessment - Estimating Severity for Filter Integrity Testing

“To FIT or not to FIT, That is the Question” , Biopharm International, Nov 2009 pp39-44

Impact of microbiological contamination Routes of contamination in the process Risk assessment and mitigation strategies Filter categorization Moderately critical filters and risk approach Critical filters and risk approach

Filter qualification

8 Elements of Sterile Filtration Qualification

Represent “worst case” process conditions, process fluid, filter performance and microbiological challenge Prove the filter’s bacterial retention capabilities with a nondestructive test.

Prove the filter removes bacteria from the stream compliant with ASTM 838-05 and regulations

Integrity Testing

Retention

Binding

Prove the filter does not unacceptably remove stream components.

Duty Prove the filter meets

QS, VMP &

all performance & duty requirements within product & process conditions.

Documentation

Sterilization

Compatibility

Prove the stream does not adversely impact the filter duty or process stream

Extractables & Leachables Identify, quantify, and assess impact of compounds that migrate from filter to process stream.

Prove the sterilization method is effective and does not compromise the filter.

Requirements for Filter Documentation Suitability for duty Process definitions Bacterial / particulate retention

Integrity testing Sterilisation process validation Adsorption

Leachables / Extractables Risk analysis approach to processing and product impact Quality by design

GMP ?

Sterile Liquid Filter Qualification & Validation – PDA Recommendations

PDA TR26 Technical report No.26, Revised 2008, Supplement Volume 62 No. S-5, Sterilizing Filtration of Liquids

Moderately Critical & Critical Liquid Qualification Differences ? Filter Qualification Element

Moderately Critical Liquid Filter Need

Duty

Chemical Compatibility

Extractables and Leachables Binding / Adsorption

Integrity Testing Sterilization

Retention

Source

Critical Liquid Filter Comment

Y

PS or UT or VT

Combination of testing and paper

Y

PS or UT or VT

Combination of testing and paper

Y

VD or UT or VT

Flush curve

TBD

PS UT VT

Not required for vessel – vessel transfer

TBD

UT VT

Based on risk

Y

UT

TBD

VD

Based on risk

Need

Source

Comment

Y

PS or UT or VT

Combination of testing and paper

Y

PS or UT or VT

Combination of testing and paper

Y

VG or UT or VT

Flush curve and TTC – based on risk

Y

PS or UT or VT

Required for pointof-fill filter

Y

UT or VT

Based on regulation

Y

UT

Y

VT

UT = Filter User Test, PS = Paper Source, VD = Vendor Document, VT = Vendor Test

Test using user fluid & conditions

Example – Buffer Filter Qualification

Duty

Yes – Paper source

Chemical Compatibility

Yes – Paper source or simple test

Extractables and Leachables

TOC, pH, conductivity flush curve

Binding / Adsorption

Document support

Integrity Testing

Post-use

Sterilization

Application dependent

Retention

Documentation support

62

Sterile Gas Filter Qualification & Validation – PDA Recommendations

PDA TR40 Technical report No.40, January/February 2005 Supplement Volume 58 No. S-1

Moderately Critical & Critical Gas Qualification Differences ? Filter Qualification Element

Moderately Critical Gas Filter Need

Duty

Source Y

Chemical Compatibility

Y

Critical Gas Filter Comment

Need

VD

Vendor doc sizing Check oxidation life

Y

PS or VD

MOC and FIT fluids comparison

Y

Source

Comment

VD

Vendor doc sizing Check oxidation life

PS or VD

MOC and FIT fluids comparison

Extractables and Leachables

N

No product contact

N

No product contact

Binding / Adsorption

N

No product contact

N

No product contact

Integrity Testing

Sterilization

Retention

TBD

Y N

UT

Vendor doc. Risk of non-integral filter

UT

Check vendor docs then do SIP study

VD

Based on risk

Y

Y N

UT

Vendor doc. Risk of non-integral filter & regulations

UT

Check vendor docs then do SIP study

VD

Actual flow < vendor study flow

UT = Filter User Test, PS = Paper Source, VD = Vendor Document, VT = Vendor Test

Impact of microbiological contamination Routes of contamination in the process

Risk assessment and mitigation strategies Filter locations and microbiological concerns Filter categorization Moderately critical filters and risk approach Critical filters and risk approach Filter qualification

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