Sampling in an Aseptic Process

Risk Mitigation & Regulatory Compliance

Somasundaram G.

Senior Technical Consultant, Asia Pacific

Process Solution, Merck KGaA

September 23, 2020

2

Agenda

1

3

4

Introduction to Sampling – Where, What & Challenges

Key considerations for Closed & Disposable sampling options

2 Regulatory recommendations and corresponding needs – Key Drivers

Complexity and Risks of traditional sampling methods

3

Introduction to Sampling

Where, What & Challenges

4

4

– Representative Sample :

– Composition

– Non-operator dependent

– Repeatable

– Protect the Process (Contamination-free)

– Protect the Person (Contained)

– Protect the Product (Contained)

– Easy of Use

– Save Time and Cost Efficient

Introduction to sampling

What are The Requirements and Challenges?

Process

People Sample

Validated

Sampling

Procedure

Validated

Sampling

Procedure

Equipment

In today’s biopharmaceutical market, sampling is critical during every step in the manufacturing process. An

imprecise or false positive result can lead to a quarantine as well as the need to repeat the analysis.

5

Sampling plan & options

USP BioreactorCell culture

media

Cell culture

supplement

Media

filtrationClarification

BBR &

Intermediate

pool

Broad

considerations

• Bioburden levels of appropriate batches to be tested based on

statistical/risk assessment

• Inoculum expansion to first active control expansion step (at scale):

Bioburden needed

• Good practice to retain samples at each expansion operation until batch

release

If clarified harvest

is filtered into a

pooling tank.

Recommended to

test bioburden of

a few batches of

clarified harvest

before filtration

Typically

prefiltration

samples (entry

point) or after

hold times

(proliferation)

Each protein pool

should be tested

for bioburden &

endotoxin

Via routine sampling

during bioreactor run

• Routine sampling may not be needed when max

preparation & filtration durations are established

• Periodic monitoring recommended annually/after

prolonged shutdowns

What are

typically

sampled?

• Cell Viability (count)

• Bioburden, including

for continuous cell

lines

• Endotoxin (not at

inoculum expansion)

• Purity check

• Osmolality

• Metabolite

• Bioburden

• Endotoxin

• pH

• Conductivity

• Media Concentration

Endotoxin

Bioburden

TOC

• Endotoxin

• Bioburden,

including

pooled

samples (prior

to filtration)

Where to Sample

6

Sampling plan & options

DSP1 AffinityVirus

Inactivation

Aggregate

RemovalCEX AEX

Virus

Filtration

Broad

considerations

• Bioburden control: some of the unit operations may not be a closed operations / or

sterilizable

• Bioburden & endotoxin testing: WFI rinse (retentate/drain line), WFI rinse (after pre-use

sanitization), EQ buffer (after equilibration step), WFI rinse (after post-use cleaning)

• Establish microbial control of reusable items (e.g. filters, resins). Acceptance

specifications for new & unused resins depends on vendor (typically ≤100 CFU/mL)

What are

typically

sampled?

• Endotoxin

• Bioburden

• TOC

• Conductivity

• Osmolality

• Impurities

• Osmolality

• pH

• Conductivity

• Bioburden

• Endotoxin

• Impurities

• Virus

inactivation

kinetics?

• Impurities?

• Endotoxin?

• Endotoxin

• Bioburden

• TOC

• Conductivity

• Osmolality

• Impurities

• Osmolality

• Endotoxin

• Bioburden

• TOC

• Conductivity

• Osmolality

• Impurities

• Osmolality

• pH

• Conductivity

• Bioburden

• Endotoxin

• Impurities

Where to Sample

7

Sampling plan & options

DSP2 /

Prep zoneUFDF

Sterile

Filtration

Sterile Bulk/

Dispensing

Operations

Buffer MixingBuffer

Holding

Buffer

Filtration

Broad

considerations

• Bioburden control: some

of the unit operations

may not be a closed

operations / or

sterilizable

• Product purity, TOC

• pH control

• Via routine sampling

• Many configurations.

• Sampling should take

risk associated with

sample & nature of

filling operation, into

consideration

• Bioburden levels of appropriate batches to be

tested based on statistical/risk assessment

• Final DF/formulation buffers: Endotoxin testing

recommended prior to use for all batches.

• Non-filtered buffer: Recommended to be

prepared fresh/used ASAP. For >24h: Hold

time validation (bioburden & endotoxin)

needed.

What are

typically

sampled?

• Endotoxin

• Bioburden

• TOC

• CIP

validation

• Impurities

• Osmolality

• Biobur

den

• Endotoxin

• Bioburden

• pH

• Conductivity

• Buffer conc

• pH

• Conductivity

• Endotoxin

• Bioburden

• Buffer conc

Where to Sample

Aseptic Filler

Sterile Hold Tank

Vent FilterVent Filter

Bioburden

Reduction

Filter

Sterilizing

Filter

Sterilizing

Filter

Prefiltered

Formulation

Sampling pointFinal Filling

Single SGF

Sampling plan & options Where to Sample

Aseptic Filler

Sterile Hold Tank

Vent FilterVent Filter

Bioburden

Reduction

Filter

Sterilizing

Filter

Sterilizing

Filter

Vent Filter

Sterilizing

Filter

Prefiltered

Formulation

Redundant Filtration

9

How to ensure that the process is microbiologically

controlled?

• Regular sampling of equipment that can not be sterilized

• Collection of samples after cleaning and sanitization

• Collection of samples during equipment equilibration and rinsing

– BPOG (2015):

– “...containers include disposable systems that are closed to the environment“

– “Bioburden samples are recommended to be stored at 2-8oC and tested within 24 hours

of collection“

10

Setting bioburden control levels (Alert/ Action)

1

0

– BPOG (2015):

– “There are no recommended bioburden levels provided in regulatory guidelines or

compendia... Manufacturers are responsible for setting bioburden control levels...“

• Most members: 1 to 10 CFU/mL. Upstream (conducive for proliferation): Near sterile;

Downstream: ≤ 10 CFU/mL

• Control levels are set prior to product licensure. “After commercialization, periodic

review is recommended to ensure control levels reflect product performance over

time.“

• Correlate bioburden testing with results of routine environmental monitoring.

11

Regulatory recommendations and

corresponding needs –

Key Drivers

11

12

Drivers for Aseptic Process Sampling

Assess the state

of the process

• Adjust sensors

• Verify, detect,adjust parameters

Transfer materials

• Adjustment by addition

• Seeding by inoculation

Extract materials for

later assessment

• Internal investigation

• Regulatory requirements

• Secure traceability

Global

Regulatory

trends:

• Process Validation

• QbD

• PAT

13

Relevant Regulatory Agencies & Industry Associations

Regulatory Agencies /

Industry Associations

Relevant

Documentation

Food and Drug Administration cGMP guidance

World Health Organization

Annex 2 – Good manufacturing guidance for API

Annex 4 - Good Manufacturing Practices for pharmaceutical

products: main principles

European Medicines Agency GMP Annex 1 - Manufacture of Sterile Medicinal Products

International Council for Harmonization of Technical

Requirements for Pharmaceuticals for Human Use ICH Q7 – GMP guidance for API

Parenteral Drug AssociationReport #69 - Bioburden and Biofilm Management in

Pharmaceutical Operations

The Pharmaceutical Inspection Convention and

Pharmaceutical Inspection Co-operation Scheme

Guide to good manufacturing practice for medicinal products –

Part I

14

Extracts of Relevant Requirements

Recommendations Source

Contamination control&

Monitoring before bioburden reduction

• PICS/S – FDA cGMP – Part 1 §5.19 –f • Use of closed system recommended from phase 1

• WHO Annex 4• “The use of disposable sampling materials has distinct advantages”

• WHO Annex 2 - ICH Q7A - GMP guidance for API• EU GMP Annex 1 • EudraLex – Vol 4 – Part II – 2009

Operator bias elimination• WHO Annex 4• FDA • European Pharmacopeia - Guidelines for Sampling of Pharmaceutical

Products and Related Materials

Representative sample

Health & safety focus

Retained samples

• FDA cGMP Guidance for the industry investigational drugs section F. Laboratory Controls / 1. Testing

• 2 years after expiration date / completion of trial and twice the quantity necessary to perform all tests

15

Sampling Process & Precautions

WHO Annex 4: Preparation for

sampling

• “All sampling tools and

implements should be made

of inert materials and kept

scrupulously clean. […].”

• “The cleaning procedure

used for all sampling tools

and implements should be

documented and recorded.”

• “The use of disposable

sampling material has distinct

advantages”

WHO Annex 4: Sampling

Operations and Precaution

“There should be a written

procedure describing the

sampling operation. […]. It

should ensure that

representative samples are

taken in sufficient quantity for

testing in accordance with

specifications.”

WHO Annex 4: Storage and

Retention

“The container used to store a

sample should not interact with

the sampled material nor allow

contamination. […]. As a general

rule the container should be

sealed and preferably tamper-

evident.”

16

Sampling of Pharmaceutical Products and

Related Materials

• FDA cGMP for phase 1 drugs

“recommends the use of closed system to minimize the risk of contamination”

•

Guidance for the industry investigational drugs section

F. Laboratory Controls / 1. Testing

“We recommend that the sample consist of a quantity adequate to perform additional testing or

investigation if required at a later date […]. We recommend that you appropriately store and

retain the samples for at least two years […].”

• Q7A GMP guidance for manufacturing API – section C. In-process Sampling and Controls (8.3)

“In-process sampling should be conducted using procedures designed to prevent contamination

[…]. […] ensure the integrity of samples after collection.”

17

Regulatory Focus on Contamination

Control during Sampling

8. Production and in-process controls

• 8.3 In-process sampling and controls

• 8.35 “In-process sampling should be

conducted using procedures designed to

prevent contamination of the sampled

material and other intermediates or APIs.

Procedures should be established to

ensure the integrity of samples after

collection.”

Annex 2, WHO GMP for active pharmaceutical

ingredients

EudraLex, Volume 4

ICH Q7A

Identical script in these 3 regulatory documents

18

8.94 Bioburden samples should be taken from the bulk product and

immediately prior to the finalsterile filtration. Systems for taking samples should be designed so as not to introduce contamination.

9.7 Sampling methods should not pose a risk of contamination to the manufacturing operations.

7.3 Non-essential processes such as product inspection

and in process testing should be conducted à with the crimping option

there is a validated and easy disconnection in place to further

treat the sampleoutside the clean areas wherever

possible.

Annex 1 EU GMP (Draft)Regulatory requirements

19

Complexity & Risk of

Traditional Sampling Methods

2020

Open Sampling Valve

Steam in place valve

Septum sampling Aseptic Connectors Tube Welding

Large number of

samples

Low cost per sample

Large number of samples

Closed sampling

Low cost per sample

Large number of

samples

Low cost per sample

Flexible & Reliable

Aseptic sampling

Safe and disposable

Large number of samples

Aseptic sampling

Flexible

Dead-leg

Loss of product (flush)

Open sampling

Impossible to sterilize

Complex operation (SIP)

Safety hazard (heat)

Risk of sample dilution

Container limitations

Not steam sterilizable

Safety hazard (needles)

Sample volume

limitation

Extra cost

Potential dead-leg

(tubing)

Limited disconnection

Waste of product

Requires utility

Piece of hardware

High risk of

contamination

Operator & Process

safety

Sample

representativeness

Operator training & safety

Sample

representativeness

High risk of

contamination

Operator safety

Sample

representativeness

Operator trainingRequires maintenance

Operator training

What are the Ways to Sample?Traditional sampling: Pros, Cons and Limitations

21

Complexity and risks of traditional sampling methods

• FDA Warning letters (483) issued in

2015

• 104 – Procedures designed to prevent

microbiological contamination of sterile

drug products not established, written

or followed

• 24 – Representative samples not

obtained

Microbial contaminations during manufacturing

• Increases risk

• Results in enormous cost

• Requires complicated investigations to

prevent reoccurrence

Cost of sampling VERSUS cost of a shut down

22

30Percent

1-6Months

1-14Million Euro

Percent of process deviations

caused by contamination*

Length of time to complete an investigation

Operations cost

*Source Langer 2013, Wiebe 2014

Biologics in-process contamination

23

30Percent

1-6Months

1-14Million Euro

*Source: Langer 2013, Wiebe 2014

Biologics in-process contamination

Impact Productivity losses

Material replacement costs

Batch loss

Interruption of product supply

Delay in clinical development

24

Key considerations for Closed

& Disposable sampling

options

25

Considerations for adopting single-use

Aseptic sampling

Situations Implications

Contamination of samples

(out of in process

specifications)

• Product batch on-hold

• Internal investigations

• Additional tests on final product

• Conflict between manufacturing and QAQC

• Risk of remarks from regulatory inspectorsContamination of process

Complex sampling procedure

• Risk of false results

• Frequent operator training

• Unhappy operators

• Internal investigations

26

Considerations for adopting single-use

Aseptic sampling

Indirect

cost

Direct

cost

• Labor costs

• Process/ product

downtime

• Scrap of batch due to

contamination

• Investigations for

product/ process

deviations

• Purchase price

• Taxes

• Freight

• Inventory costs

Total cost of

ownership

(TCO)

27

Self-assessment: Am I suitable to adopt single-use

Aseptic sampling?

Considerations Considerations

1 2 3 1 2 3

It is difficult to implement changes to

my infrastructure (e.g. piping).

There is high risk of bio-

contamination in my process.

There are challenges to conduct

cleaning validation.

My current sampling method has

risk of cross-contamination.

My operators are very familiar with

plastic components in the process.

I need to collect samples anywhere.

I have limited process time to

prepare sampling assemblies

(traditional).

I need to collect large numbers of

samples.

I have limited cycle time. I need

quick turnaround between batches.

I require a varied range of sampling

containers.

I am concerned about operator

safety.

My process is sensitive to price.

I have limited resources for operator

training.

Strong

YES

Strong

NOStrong

YES

Strong

NO

28

Sterile sampling value assessmentCase study example

62% Reduced labor hours

35% Reduced product loss during

sampling

80% Reduced deviation costs

400 Autoclave cycles eliminated10%Overall cost reduction

Key facts

Background

Replace standard open sampling with NovaSeptum® sterile sampling assembly

Outcome

29

• There are complexity and heightened risks in using traditional

sampling methods.

• Regulatory recommendations are in place and single-use aseptic

sampling could help meet several corresponding needs.

• There are several key considerations to design/optimize your sampling

plan.

Conclusions

30

Acknowledgements

Mark Antoine Kaag, Global Product Manager, France

Janmeet Anant, Global Regulatory Advocate, USA

Thank You