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Recommended Practices for Assuring

Integrity of Single-Use Systems

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The Bio-Process Systems Alliance

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T
he increasing uptake of single-use between suppliers and end users to

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technologies (SUTs) in critical achieve the critical objectives. A risk-
current good manufacturing management plan for single-use

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practice (CGMP) processes and systems (Figure 1) can help companies
applications has made their integrity a address potential risks both in system
critical quality attribute (CQA) for both manufacturing and in end-user

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suppliers and end users of such systems. processes. Failure mode and effects

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Current regulations focus on final analysis (FMEA) typically addresses
packaging, however, without taking into supplier steps such as assembly, testing,
account the unique aspects of assemblies M packaging, and shipping of empty
used in bioproduction. Ongoing components and assemblies; and user
initiatives include revision of PDA TR 27 steps such as unpacking, handling,
(1) and creation of ASTM workstreams (2, installation, and processing.
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3) to propose good practices for the Quality by design (QbD) provides
Working with single-use bag assemblies
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integrity of single-use systems. In the another risk-management tool for

absence of a regulatory framework or stage through manufacture, assembly, integrity assurance. It demands an
P

guideline specifically applicable to such packaging, sterilization, validation, understanding of the risks and
systems used in bioproduction, a risk- certification, and shipping of systems to potential defects associated with each
management approach is recommended. end users. Suppliers also help end users stage of the SUT life cycle (Figure 2),
H

In addition, scientific understanding of in areas such as application studies, starting with suppliers, from design
the critical defect size for a risk of liquid operator training, and procedures to and development stages through to
IT

leakages and/or microbial contamination verify and maintain system integrity assembly, manufacture, validation, and
is a prerequisite for integrity control both before and after use. packaging. After product irradiation
W

strategies to correlate maximum That shared responsibility demands and shipment, end users become
allowable leakage limits (MALLs) with collaboration and close partnership responsible for implementation,
the detection limits of physical testing
that may be applied to ensure the Figure 1: Risk management of single-use systems
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microbial integrity. According to

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USP<1207>, the MALL is the greatest Lifecycle Visual inspection

leakage rate (or leak size) tolerable for a management Integrity testing
Cost–benefit QbD approach
given product/package that will pose no
analysis
R

risk to product safety or inconsequential Leak testing

impact on product quality (4). Business Integrity
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RisK-Based Approach to
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Assessing System Integrity

For multiuse systems, end users hold Process
R

Transport/storage
primary responsibility and control over SAL 10–6
Operator and Operator training
system design, construction, integrity, Barrier properties Product
Environment Assembly/disposal
operation, and maintenance. With Control strategies
Containment strategy
single-use systems, by contrast, Materials science
Health, safety,
suppliers/integrators take increased Suppliers, end users and environment
responsibility from the earliest design Verification/validation

30 BioProcess International 17(3) March 2019

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Figure 2: Single-use system product lifecycle
Practical Testing for Assurance of
Integrity: Leak detection in single-use
Supplier Integration systems may be possible through visual
SU critical inspection for major flaws that generate
component gross leaks. In most cases, the location
Assembly of such flaws may be indiscernible or
Packaging
the holes too small for detection by
Other human eyes. Conversely, visual
component
inspection often turns up findings that
Irradiation are not actually defects. Practical
testing would alleviate such false
findings. More sensitive detection
methods are essential to mitigate risks
End-User Implementation such as microbial contamination. Two
types of nondestructive tests meet that
Disposal Use Installation Shipment requirement and thus are suitable for
single-use systems: those based on air-
pressure and trace-gas measurement.
Pressure-Based Tests: The basic
principle of a pressure test is to detect
operator training, assembly provide the level of sensitivity required leaks in a system by inflation of its
installation, use, and disposal. to confirm that a total barrier against components with air to a defined
Practical controls such as visual microbial ingress or liquid loss exists. pressure. The flow of that air through
inspection and integrity testing are defects can be detected either by
important considerations in Collaboration Between pressure decay or direct flow
manufacturing validation for Suppliers and End Users measurement. Both approaches depend
components and single-use systems. Depending on the criticality of the unit on the ideal gas law: PV = nRT, where
These procedures normally should be operation in which a single-use system p = pressure, V = volume, n = the molar
included in each supplier’s actually gets implemented, value, R is the gas constant, and
manufacturing controls. But when it is requirements for validation and testing T = temperature. Higher pressures and
technically feasible, in-process are likely to differ. For unit operations lower test volumes enhance sensitivity,
implementation of leak testing and considered to be high risk for breaches and maintaining a constant
integrity testing can be valuable to end of integrity, additional testing during temperature is necessary throughout
users as well. It may be unacceptable, system manufacturing might be the test.
however, if such testing adds undesirable required. Table 1 provides some Direct air-flow measurement monitors
complexity to systems and standard guidance for supplier evaluations during the gas-flow rate required to maintain
operating procedures (SOPs). User testing different stages of SUT development, system pressure during testing. The
even can introduce a risk of a false- validation, manufacturing, and main differences between this and the
failure results. For large and/or complex transportation. And Table 2 summarizes pressure decay method are that
single-use systems, current integrity test end-user assurance of integrity during • the air supply is not isolated during
methods may provide a valuable test the stages of SUT use in drug the test
method for gross defects but be unable to manufacturing. • test pressure is held constant

Table 1: Potential testing/qualification approach performed by suppliers of single-use systems for integrity assurance

Single-Use System Single-Use System Single-Use System

Development and Validation Manufacturing Transportation
QbD, Risk Assessment, Statistical Process Control
Unit Operation Criticality Process Validation and QC Testing Design and Packaging
LOW RISK: • Component validation • Component testing (e.g., filters, sterile • Packaging validation
Upstream • Mechanical tests connectors, bags) • ASTM/ISTA transportation
Sterile filtration possible • Assembly validation  Junction test • Seal quality tests (e.g., bags) validation
• Shelf life • Visual inspections of system assembly
MEDIUM RISK: Bulleted items above Bulleted items above and Bulleted items above
Downstream (including final • Leak testing of bags depending on joint risk
formulation and compounding) assessment
Sterile filtration possible
HIGH RISK: Bulleted items above and Bulleted items above and Bulleted items above
Filling • Microbial aerosol or immersion • Integrity testing of system assembly with
No sterile filtration possible challenge test sensitive test method (e.g., helium, gas tracer)

32 BioProcess International 17(3) March 2019

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Table 2: Potential end-user strategy to ensure integrity throughout the life of single-use systems (SUS) based on risk assessment
End-User Single-Use Product Commercial Commercial
End-User Evaluation
Implementation Transportation Preuse Testing Postuse Testing
Design and Development Validation and Training Incoming Inspection Preuse Installation Postuse Testing
• Process mapping for intended use • User qualification package; • Checked at reception and • Visual inspection of • Visual inspection for
• Design space installation and performance intermediate storage packaging and SUS absence of liquid leaks
qualification (IQ–PQ) and location (if any) for visible • Nondestructive integrity • Sterility testing of product
• Review of existing validation and aseptic broth validation damage
gaps identification testing • Additional testing per
• Consistency batches; • Quality control (QC) • Visual checks during use: user’s requirements
• Definition of a manufacturing training by supplier and/or end- inspection
control strategy connectivity, tubing
user subject-matter expert • Documentation installation, clamps, special
• Verification of supplier validation (SME) attention to sterile
package • Visual inspection of SUS connections
• Establishment of SUS process and • Verification of integrity: for
storage conditions based on critical applications,
supplier recommendations; visual submission of SUS to full life
inspection cycle and verification of
• Integrity verification integrity at the end

• air flow through defects is aerosol challenge test is appropriate for

measured directly on the upstream side. most applications because it
In small single-use systems (e.g., flat
A quality risk management corresponds to worst-case conditions for
bags), pressure-decay and flow- strategy for integrity a single-use system. The liquid
measurement integrity tests can detect assurance of single-use immersion test is the method most often
barrier defects ≥10 µm in size. Such a systems demands an used for sterile products in their final
detection limit can be correlated to a dosage containers.
liquid leak and/or microbial ingress
understanding of their Both methods are probabilistic,
result under real-use conditions. For lifecycle and a however. They differ in sensitivity to
large-volume 3D systems, the limit will STRONG working microbial ingress, which in both
be ≥100 µm. Because defects of that size relationship between methods is highly dependent on test
can allow ingress of microbial conditions. The choice of test and test
contaminants under some process
suppliers and end users. conditions should be informed by risk
conditions, the test may be defined assessment of a given SUT application.
better as a gross leak test or with a detection limit of ≥2 µm. Such The critical defect size can be
postinstallation test. detection limits can be correlated to the different according to the type of
Trace-Gas–Based Tests: Typically most stringent immersion and aerosol challenge test used and its detailed
using helium, trace-gas integrity testing bacterial challenge tests, helping to parameters, as well as the size and
offers a suitable method for single-use assure the microbial integrity of systems shape of test organisms, differential
systems with which the highest possible tested. Applicability of the helium test pressure across defects, and so on. Other
level of sterility assurance may be method does have limitations, however, factors related to process and product
required. The principle of this method is depending on single-use system also affect the critical defect size. For
to measure the amount of a tracer gas assembly volume, complexity, materials, example, a corrosive fluid with no
leaking through barrier defect(s). A test and so on. susceptibility to bacterial infection but
container is placed inside a rigid requiring zero liquid loss for safety
chamber and connected to a helium Critical Defects and Correlation reasons might allow use of a bag with a
inlet valve. Air is evacuated from the by Microbial Challenge relatively large critical-defect size.
chamber, and helium is injected into the More sensitive tests will have limited Sterile products, however, present the
single-use bag. If barrier defects are significance if they cannot be correlated most demanding requirements for
present, the vacuum will pull helium to some form of microbial challenge system integrity. Single-use systems for
from the single-use system into the during the validation phase to assess such applications must be validated
vacuum chamber. Then helium in the the risk of microbial ingress and using appropriate microbial challenge
chamber is detected and quantified by a demonstrate container closure integrity tests to determine the smallest critical-
mass spectrometer, and the amount of (CCI). For final drug containers, the defect size for prevention of microbial
helium released can be correlated with MALL is defined as the maximum leak ingress either under worst-case
defect size. size that poses no risk to product safety conditions or those simulating typical
As of today, the helium test method (4). For each unit operation, it should be process applications. An ASTM
most suitable for single-use systems determined based on risk assessment workstream on microbial ingress testing
(including some 3D systems up to several and intended system use. of SUTs has been initiated and should
hundred liters in volume) is the most Currently, two basic tests methods provide more guidance on this specific
sensitive integrity test method available, are used for microbial challenge. The topic (3).

34 BioProcess International 17(3) March 2019

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Implementation of Integrity Tests Figure 3 lists options available for manufacturing is part of the risk
Control of integrity assurance can be implementation of such testing. mitigation strategy. Through operator
achieved only through combinating a A quality risk management strategy training and implementation of detailed
risk-based QbD approach with, when for integrity assurance of single-use SOPs, end users can ensure that
technically practicable, a selective systems demands an understanding of integrity will be maintained during
application of integrity tests and gross their lifecycle and strong working installation and operation. Based on
leak tests together with visual relationships between suppliers and end SUT application risk assessment, further
inspection. Testing may be considered at users (Figure 4). Suppliers should use assurance of integrity could be required
three stages of the SUT life cycle: QbD principles during system design and achieved by performing optional
component testing by the system and development and thoroughly tests at the point of use. Work initiated
manufacturer, assembly testing by validate their assembly, manufacturing, in ASTM workstreams will continue to
suppliers/integrators, and assembly and packaging processes. Leak and frame existing recommendations from
testing by end users at the point of use. integrity testing during the SUT the Bio-Process Systems Alliance (BPSA)
for integrity assurance of single-use
Figure 3: Integrity assurance throughout the life of single-use products systems.

Assembly Supplier/Integrator References

Validated by 1 PDA Technical Report No. 27:
component
Pharmaceutical Package Integrity. 52(S2)
manufacturer Qualified
assembly process
1998.
2 ASTM E55.04-WK64337: Best Practices
— Integrity Assurance and Testing of Single-
SU critical Use Systems. American Society for Testing
component
and Materials: West Conshohocken, PA, 13
Assembly Packaging July 2018.
Other 3 ASTM E55.04-WK64975: Testing
component Method — Microbial Ingress Testing on
Single-Use Systems. American Society for
Optional leak or integrity Testing and Materials: West Conshohocken,
testing depends on system Irradiation PA, 30 August 2018.
Optional leak or
integrity testing size, complexity, and 4 USP<1207> Package Integrity
practicality.
Evaluation: Sterile Products. United States
Pharmacopeial Convention, Inc. Rockville,
MD, 1 May 2018.

End-User Implementation
Further Reading
Vanness B, et al. Response to the
Disposal Use Installation Shipment
15(1) 2017: 20–21. c
Publication of USP ‹1207›. BioProcess Int.

Optional pre- or postuse

leak or integrity testing Operator To share this in PDF or professionally printed
depends on system size, training format, contact Jill Kaletha: jkaletha@
complexity, practicality, and mossbergco. com, 1-574-347-4211.
criticality of application.

A White Paper
Figure 4: Best practices for risk mitigation; QbD = quality by design, QC = quality control This article is a summary of the Bio-Process
Systems Alliance (BPSA) initiative from a task
force made up of industry experts from end-
user and supplier companies. The published
white paper, Design, Control, and Monitoring
of Single-Use Systems for Integrity
Assurance, can be downloaded at
Development, http://bpsalliance.org/technical-guides.
Design, Validation Manufacturing User

QbD/risk assessment;
QbD/risk assessment; QC, visual control, operator training, QC,
mechanical, microbial, documentation, visual inspection,
shipping and integrity optional supplier optional point-of-use
tests; SUS fit for purpose leak/integrity test leak or integrity test

36 BioProcess International 17(3) March 2019

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