TECHNICAL ASEPTIC

VALIDATION OF ASEPTIC
PROCESSES
Using Media Fill
By Richard Chai and David J. W. Barber, PhD, CBiol, MRSB, PCQI

Aseptic process simulation (APS) is essential analysis, and further media simulations may be required to com-
plete the validation.
for validation of an aseptic manufacturing
Aseptic processes are typically carried out in conventional
process and is required by regulators to cleanrooms with vial fi lling and stoppering in Grade A laminar
demonstrate the aseptic capability of such airflow (LAF) in a Grade B background environment. The fi lling
processes. A successful program of APS and environment may be further protected within a restricted-access
barrier system (RABS) with glove ports for access to the filling line.
aseptic manufacturing requires significant
Alternatively, processing equipment for the critical steps may be
operator training, skills, and supervision; enclosed in a glove box or isolator. Each of these systems enhances
thorough maintenance; effective cleaning and the filling environment’s sterility assurance but also presents
disinfection; significant oversight of every aspect challenges for material transfer, operator access, environmental
monitoring, and APS.
of the operation by quality assurance; and
microbiological monitoring by quality control. REGULATORY EXPECTATIONS

A
Aseptic manufacturing and validation follow current GMPs and
n overall validation of aseptic processing (as distinct from related GMP Annexes and Guidance. These pertain to the manu-
manufacturing process validation [PV]) is used to assess the facture, validation (APS), and control of sterile products for
contamination risk of an aseptic production process by injection (as well as eye drops and advanced therapy medicinal
simulating the manufacturing process using microbiologi- products). Current guidelines come from the European Union/
cal growth media instead of the drug solution. This is necessary in Pharmaceutical Inspection Convention (EU/PICS), China (2010)
part because the sterility test used to release batches of sterile GMP (NMPA), United States Food & Drug Administration (US
products has inherent limitations in detecting contaminated FDA), and World Health Organization (WHO) [2–13]. They may
units in batches with low levels of microbial contamination, due to reference related International Organization for Standardization
the limited number of samples that can be removed for destructive (ISO) and Parenteral Drug Association (PDA) standards [14–17],
testing; this relationship has been evaluated statistically [1]. such as those relating to cleanroom air-cleanliness classification
Sterility assurance in aseptic processing requires contributing and particle monitoring [17].
elements—such as the heating, ventilation, and air conditioning Because of the high safety risk profile for parenteral drug
(HVAC) system, cleanroom environment, material transfer, equip- products, the protocols, results, and reports for APS form an inte-
ment, and manufacturing process steps, including sterilization gral part of regulatory submissions for such products, meaning
processes and sterilizing fi ltration—to be qualified and validated they are included in investigational new drug (IND) applications,
as applicable and for personnel to be trained and qualified. new drug applications (NDAs), and marketing authorizations
Simulation of aseptic manufacturing processes using liquid (MAs). Ancillary documents such as training records, environ-
microbiological growth medium (also referred to as media simula- mental monitoring reports, deviations, and investigations are key
tion or APS) is required by regulators to demonstrate the aseptic topics of scrutiny during facility inspections, as well as the qualifi-
capability of these processes. cation of facility, the equipment and utilities, and the process
APS consists of three consecutive media simulations with validation.
designated personnel in the specific cleanroom environment, fol- The expectation in APS is twofold. First, it must achieve three
lowed by repeat media simulations at six monthly intervals. Any consecutive media batches that meet target acceptance criteria.
media fi ll failures require thorough investigation and root cause Second, the solution filtration process must be validated against a

50 P h a r m a ce u t ic a l E ngine e r ing
microbial challenge with 10 7 colony-forming units per square Figure 1: Points to consider when designing the media fill study.
centimeter of fi lter medium (using Brevundimonas diminuta, a
small-celled Gram-negative bacterium suspended in the drug
solution).
Examples of media fill run sizes and acceptance criteria for
APS that have been incorporated in GMP Annex [6] and Guidance Worst-case
[10, 11] include: challenge

u When fi lling less than 5,000 units, zero contaminated units Records and
microorganism
should be detected. A contaminated unit is considered cause identification Routine/
and nonroutine
for revalidation following an investigation. Operator training interventions
u When filling 5,000 to 10,000 units, one contaminated unit and qualification
monitoring
should lead to an investigation, including consideration of a
repeat media fi ll. Following investigation, two or more con- APS
taminated units are cause for revalidation. (media fill)
u When fi lling more than 10,000 units, one contaminated unit
should lead to an investigation, and two or more contami- Unit handling, Lyophilization
nated units are cause for revalidation. incubation, and (if applicable)
inspection

APS CONSIDERATIONS
The following is an overview of points to consider when designing Microbiological
the media fill study for an aseptic manufacturing process. growth medium

Worst-Case Challenge
APS should mimic, as closely as possible, all aspects of the aseptic
manufacturing process and should involve a “worst-case”
approach as a challenge to the robustness of the aseptic operations.
The “worst-case” should be defined with supporting rationale. should be compliant with current GMPs, and APS must not be used
Risk assessment principles should be used to determine the to justify poor aseptic practice or equipment design.
worst-case challenges related to line speed, container size, batch Routine interventions include charging stopper and seal hop-
size, hold time, configurations, and operating conditions. pers, removing jammed stoppers or toppled vials, taking environ-
Some examples of worst-case challenges : mental monitoring samples (settle plates, active air samples, and
u Filling process contact plates), and checking in-process control samples (e.g., man-
u Aseptic assembly of equipment and aseptic connections ual weight checks). Routine interventions should be performed as
prior to commencement of fi lling described in the production standard operating procedure (SOP) or
u Slowes t f i l l i ng s pe e d w it h w ides t open i ng v i a l s/ the batch record or environmental monitoring SOP. Procedures to
containers be followed in the event of machine jams and spills may include
u Maximum filling volume for small vials/containers, partial line clearances, including removal of exposed units.
due to ha nd l i ng d i f f ic u lt y t hat ca n resu lt i n more Nonroutine interventions may include changing the filling
interventions nozzles or handling unexpected events, such as breakdown main-
u Maximum batch fi lling duration (may include lyophilizer tenance, line stoppages, machine adjustments, and material
loading and door opening duration) transfers. Interventions can also be grouped by access point, and
u Operator fatigue as contamination risk their risk assessed so that worst-case (highest risk) interventions
u Operating conditions are included in the study.
u Maximum number of personnel in aseptic area
u Shift changes, personnel changes, and operator breaks Lyophilization
u Hold time EudraLex Annex 1 (2009) [6] states, “The process simulation test
u Equipment/room clean hold time should imitate as closely as possible the routine aseptic manufac-
u Equipment sterilization hold time turing process....” It is unlikely that the exact lyophilization cycle
for the product can be replicated during media simulations due to
Routine and Nonroutine Interventions the constraint of maintaining the media to support microbial
Interventions to be included for simulation in the media fill proto- growth. Deviation from the production cycle must be justified. For
col include routine and nonroutine manipulations by operators. example, if the recommended temperature range for media is 5°C
The regulatory expectation is that interventions included in APS to 25°C, the chamber pressure, normally 100 to 200 mbar, should

m a rch /a p r il 2 0 2 2 51
 TECHNICAL ASEPTIC

not be lower than the equilibrium vapor pressure of the media at Unit Handling, Incubation, and Inspection
the loading temperature to avoid boiling away the media and to After fi lling, stoppering, and sealing, 100% visual inspection is
avoid overconcentration of media, which could adversely affect performed for defects such as the presence of visible foreign
the recovery and growth of microorganisms. matter, high or low fi ll volumes, and damaged vials, stoppers, or
The chamber dwell time during APS does not impact risk seals. Such defective units would be normally removed (rejected)
because the higher chamber pressure required to avoid boiling of from product batches, but in the case of APS batches, such defec-
media does not require the use of a pressure control (gas injection) tive integral units must be retained and all such containers must
system. In the absence of airflow transport mechanism and turbu- be incubated. If fi lled containers are broken or otherwise dam-
lence, the chamber dwell time becomes immaterial during APS. aged so that they are nonintegral and potentially contaminated,
Based on risk analysis, the aeration or vacuum-break step in the they must be recorded and reconciled with the batch record
lyophilization cycle may have higher risk of contamination because quantities. All appropriate media fill container units must be
it involves air turbulence [18] and the possibility of entrained parti- incubated.
cles entering the containers. Because the application of full vacuum The incubation conditions selected are optimal for recovery
is not possible during APS, multiple partial vacuum steps should be and to allow for detection of both slow-growing and normal con-
considered to simulate the worst-case aeration. The media volume taminating organisms, i.e., adequate to detect microorganisms
in the vials before lyophilization must ensure the wetted surface of that might otherwise be difficult to culture. The incubation condi-
the container mimics the production case. tions used generally are 20°C to 25°C for seven days (lower temper-
Media simulation of the lyophilization step could involve ature first) followed by 30°C to 35°C for a further seven days.
loading the required number of media-fi lled vials as per the rou- Containers are typically incubated on their sides, and while
tine commercial production procedures, while assuring the time subjected to each incubation temperature, turned at least once to
that the door is open to the cleanroom environment is at least as ensure that the entire interior surfaces of the vials and the stop-
long as the maximum time incurred when loading a commercial pers are contacted by the growth medium.
batch of product. Records (chart printouts or electronic records) of the incuba-
Once the modified media lyophilization cycle has been com- tion conditions must be maintained, including the date and time
pleted, the chamber vacuum should be broken using sterile- of incubation commencement, turning of vials, transfer to the
fi ltered compressed air so that all units are stoppered under pres- second incubator, and further turning and completion of incuba-
sure to avoid inhibiting microbial recovery and growth. (Sterile- tion. Incubated vials must be inspected by operators qualified to
filtered nitrogen gas should not be used to break the vacuum distinguish sterile vials (“no growth”) from vials showing micro-
unless a specific anaerobic media simulation is undertaken.) bial growth (surface pellicle or turbidity in the solution). A small
number of sterile (“no growth”) vials should be selected from the
MICROBIOLOGICAL GROWTH MEDIUM incubated vials for use as after-test growth controls; these vials are
Media for microbiological recovery and growth are defined in then inoculated with ≤ 100 colony-forming units of the compen-
pharmacopoeia—such as the United States (USP), European (Ph. dial microorganism strains mentioned previously, and incubated,
Eur.), Chinese (ChP), and Japanese (JP) Pharmacopoeia—and followed by inspection for positive microbial growth.
should be made and sterilized according to the manufacturer’s
instructions. The media used in APS for filling sterile, depyrogen- Environmental Monitoring
ated containers is generally tryptone soya broth (TSB), or soybean During APS, all routine and normal processes (such as cleaning,
casein digest medium (SCM), which supports recovery and growth disinfection, and maintenance) should be continued to maintain
of viable aerobic microorganisms. Anaerobic growth medium the cleanroom environment in qualified status. This includes
such as fluid thioglycolate medium (FTM), which supports recov- particulate and microbiological environmental monitoring,
ery and growth of obligate or facultative anaerobic bacteria, may which can demonstrate that the specified cleanroom environment
be considered under special circumstances (e.g., where the product conditions are maintained. These monitoring results may provide
solution is to be filled into nitrogen-flushed vials). key information for the investigation of a failed media run.
The growth medium, supplied as a dry powder, is a critical Particulate monitoring during aseptic product filling and APS
material for APS. It is recommended that the manufacturer is consists of continuous monitoring for particulates in the < 0.5 μm
qualified and monitored as an approved supplier; a growth promo- and < 5.0 μm ranges, using a particle sampler attached to an isoki-
tion certificate may be obtained with every batch. Prior to release netic probe located near to the point of fi ll in the Grade A area. A
for use, batches of the media to be used for APS should be reconsti- permanent record of the particle counter’s printout (or certified
tuted and sterilized; then samples should be subjected to quality true copy if the printout is on thermal paper) must be attached to
control testing for growth promotion by inoculating with ≤ 100 the batch record for the product fill or APS batch. The regulatory/
colony-forming units of representative compendial strains of action limits for the monitoring, per m3 air volume, are not more
microorganisms. Microorganism strains from environmental than 3,520 particles in the < 0.5 μm particle size range and not
monitoring may be included in the growth promotion test. more than 20 particles in the < 5.0 μm range.

52 P h a r m a ce u t ic a l E ngine e r ing
 The microbiological methods used should be described in an
SOP, including a map of the locations at which the samples are to be
taken or plates exposed. Each batch of environmental sampling
plates must be tested for sterility and growth promotion capability
against the recommended compendial strains of microorganisms
before release for use. APS consists of three
The methods used for environmental monitoring are stated in
China GMP [3] and EudraLex, current Annex 1 [6]: active air sam- consecutive media simulations
pling (1 m3 sample volume) onto 90 mm agar plates; settling plates
90 mm in diameter, with exposure up to 4 hours (if the APS or
with designated personnel
production filling lasts longer, new settling plates must be exposed
for each subsequent 4-hour period); surface contact plates 55 mm
in the specific cleanroom
in diameter (in which the plates are contacted against machine
surfaces or cleanroom walls, floors, or operator gowns); gloved-
environment, followed by
finger samples performed by cleanroom operators during the fi ll- repeat media simulations at
ing period and upon leaving the cleanroom, taken by contacting
four fingers and thumb onto the surface of a 90 mm tryptone soya six monthly intervals.
agar (TSA) settle plate.
Media is usually TSA for viable aerobes or sabaroud dextrose
agar (SDA) for fungi (molds) and yeasts. Surface contact plates may
be TSA, usually incorporating a neutralizing agent to counter
detergent residues from the sampled surfaces. Agar residues are
removed from the sampling locations by wiping with 70%
alcohol. Operator Training and Qualification
The expected (regulatory) action limits for the microbiological Prior to APS batch manufacture, operators performing APS must
monitoring results of the Grade A cleanroom areas (Grade A LAF be trained in relevant procedures, including cleanroom gowning,
in Grade B background; RABS; isolator), including during APS, in aseptic connections, and correct cleanroom behavior, as well as in
colony-forming units are tabulated in China GMP [3] and EudraLex, product-specific manufacturing procedures. All staff qualified to
current Annex 1 [6]. Adjacent Grade B, C, or D cleanrooms through work in the area, including maintenance personnel, need to be
which operator gowning and material transfer for the APS occur included in APS.
should also be monitored; the stated regulatory (action) limits for Relevant training points:
these cleanroom grades are also included in the China GMP [3] and u Sterile materials and equipment should be handled only with
EudraLex, current Annex 1 [6]. The frequency of monitoring Grade C sterile instruments, such as forceps. Between uses, instru-
and D cleanrooms is to be determined based on quality risk assess- ments should be protected from contamination.
ment because such monitoring at the time of an APS may help u After initial gowning, sterile gloves should be regularly sani-
investigate any discrepancy or failure. tized by spraying with a qualified sanitizing agent such as
sterile 70% isopropyl alcohol (IPA) to minimize the risk of
Records and Microorganism Identification contamination. Personnel should not directly contact sterile
In APS batches, the numbers of colony-forming units recorded products, containers, components, or critical surfaces.
on the environmental monitoring plates in Grade A (LAF, RABS, u Rapid movements create turbulence in the critical environ-
or isolator) and Grade B areas should be recorded. An isolate ment, disturbing LAF and the integrity of sterile environ-
should be taken from each visually distinct microbial colony ments, and entraining particles. Operator movements should
and identified by species using available biochemical and/or be slow and deliberate.
nucleic acid identification methods so it can be compared with u Aseptic operators should not disrupt LAF designed to protect
organisms in contaminated units that arise during the APS. critical surfaces. When performing aseptic manipulations
This information will be critical in investigating and determin- (such as making aseptic connections, removing samples, or
ing corrective actions in the event of an APS media fill that retrieving fallen or jammed components from a fi lling line),
exceeds acceptance criteria. Environmental samples (those operators should be trained to approach the location slowly
with colonies) from Grade C and D cleanrooms should be enu- and deliberately from the side whenever possible.
merated and preferably also identified, as the information u After initial theoretical training, aseptic training operators
regarding the numbers, species, and locations of contaminating should be allowed to practice their movements in a mock-up
microorganisms may prove crucial in the investigation and or nonsterile practice environment before being permitted to
resolution of a failed media fi ll. participate in operations in the cleanroom environment.

m a rch /a p r il 2 0 2 2 53
 TECHNICAL ASEPTIC

Figure 2: Media fill failure root cause investigation and identification using an Ishikawa diagram.

Materials Environment Operator

Sterility issues Inadequate bioburden control Lack of aseptic behavior

Airflow pattern/pressure
Nonintegral containers differential issues Lack of aseptic technique

Damaged gloves, Failure to comply

HEPA filter leakage
cleanroom garments with procedure
Media Fill
Failure
Inadequate laboratory Equipment design does not
process/procedures facilitate aseptic technique
Inappropriate/high-risk Lack of sanitary design in
interventions process equipment
Incubated nonintegral Process equipment
vials/containers cleaning not robust

Method Machine

MEDIA FILL FAILURES AND ROOT CAUSE DETERMINATION organism in the failed media units, or a significant processing
A key step in the investigation is identifying microorganism(s) discrepancy or error or equipment failure.
species in positive media vials and any colonies appearing on
environmental monitoring plates, particularly those from the Case Study
Grade A/B environments, including from RABS/isolator monitor- In a sterile injectables manufacturing plant, a routine media fi ll
ing. Identification of species from colonies on plates exposed in the showed growth in one vial. The microorganism was a micrococ-
lower-grade adjacent cleanrooms, through which materials or cus, typically associated with human skin, attributed to an engi-
personnel have accessed the filling rooms, may also be crucial. neering intervention using an unsterilized tool and not reflective
The review of the deviation should encompass the preparation of normal practice. A repeat media fill was done, which also
and manufacturing processes—including cleanroom cleaning showed g row t h i n one v ia l w it h no obv iou s root cau se.
and disinfection, components and materials sanitization/sterili- Manufacturing of product was put on hold. Following an investi-
zation and transfer processes, HVAC and cleanroom operating gation, it was noted that the APS included approximately 80
parameters during the fi lling period, fi ltration process and integ- interventions to simulate any possible activities that might be
rity tests, filling operation, stoppering and capping equipment, required in normal production. However, in normal production,
and taking and transferring in-process or environmental samples. far fewer (< 20) interventions occur routinely. Therefore, it was
The review should focus on documentation, including any devia- concluded that the process may have been excessively stressed and
tions or atypical events, but may also include a review of CCTV was not representative of the commercial process being simulated.
records of the filling rooms and operations and documented Three further media fi lls were initiated, of which the fi rst media
interviews with operators. Review should also include recent fill showed growth in one vial.
engineering work or prior media fill batches. The investigation using RNA ribotyping identified that the
An Ishikawa diagram showing cause-and-effect links to a spe- microorganism in all three media fi lls showing growth was the
cific failure is a useful tool that can be used to investigate and same—a micrococcus. Microbial testing showed that one operator
identify the root cause of a media fill failure (see Figure 2). tended to shed greater numbers of skin particles than other opera-
Based on the potential root cause interactions identified in tors, including this microorganism. The investigation also identi-
Figure 2, investigating the possible failure modes and correspond- fied variability in how materials were passed into the sterile core,
ing risk mitigation measures will be necessary (Table 1). potentially providing a route of ingress.
In the investigation, different possibilities may provide the A risk assessment was carried out to determine any safety
evidence to support root cause determination, such as the ability issues arising from the sporadic low-level contamination in the
to match the identification of an environmental isolate from the process. It was concluded that based on the nature of the microor-
current (or recent) batch with the identity of the contaminating ganism, the sterility assurance levels achieved by the process, and

54 P h a r m a ce u t ic a l E ngine e r ing
Table 1: Potential causes of media fill failures.

Description Possible Failure Mode Risk Mitigation

• Excessive and unnecessary touching of surfaces

Aseptic behavior
• Talking unnecessarily in critical areas
• Inadequate sanitization of gloved hands/surfaces after high-risk • Periodic aseptic behavior and aseptic technique training
activities
Operator

• Periodic audit on aseptic behavior and technique

Aseptic technique • Rapid movement in critical areas
• Activities that may compromise sterility, such as actions above
sterile open vials/containers
Compliance to procedure Tasks not performed according to procedure Periodic audit to ensure strict adherence to procedure

Equipment design not facilitating aseptic interventions, and Design qualification; modifications may be required to mitigate the risk
Equipment design
increased risk of contamination during intervention

Inadequate sanitary design in process equipment leading to

Sanitary design
Machine

cleanability issues
Design a robust cleaning procedure considering the type of soil, cleaning
Process equipment cleaning procedure not robust
parameters, and the use of appropriate cleaning detergent, if required
Process equipment cleaning
Reduced cleanability of stainless-steel surfaces due to corrosion,
Periodic maintenance of stainless-steel surfaces to minimize risk of corrosion
which could lead to formation of biofilm
Inadequate cleaning and disinfection program for cleanroom Design a robust cleaning and disinfection program using sanitizers,
Bioburden control
surfaces disinfectants, and sporicides
Environment

Airflow pattern/pressure • Inappropriate airflow pattern in critical area • Remediate airflow pattern to minimize risk of contamination to products
differential • Differential pressure excursions in critical room/area • Ensure interlocking of doors
Leakage Leakage in HEPA filter Periodic maintenance and leak tests

Laboratory process/procedures Laboratory process/procedures inadequate Review and remediate laboratory procedures to minimize errors

Inappropriate/high-risk High-risk interventions disrupt unidirectional (laminar) airflow, and Smoke studies to be conducted and evaluated for risk of contamination
Method

interventions thus increase the risk of contamination (turbulence) for each intervention
Incubated nonintegral vials/
Failure to spot nonintegral vials/containers Training of inspectors on nonintegral vials/containers
containers

Parts/packaging components not sterile due to sterilization process Verify sterilization processes meet acceptance criteria and assess impact of
issues any excursions
Sterility issues
Sterility of parts/packaging components compromised after Ensure sterile parts/packaging components are protected from contamination
Material

sterilization, prior to usage through the use of appropriate protective barrier

Nonintegral containers Nonintegral vials/containers not segregated Training on inspection of nonintegral vials/containers prior to incubation
Damaged gloves, cleanroom
Damaged gowns and gloves can increase risk of contamination Confirm the integrity of the gowns and gloves visually
garments

the regulatory guidelines, the safety risk was low. However, it was identified during the investigation—reallocation to other duties of
now obvious that the process was not operating in a validated the “shedding” operator and reduction in number of interventions
state. No further batches of the product were manufactured until simulated per media fill (the interventions were divided into three
the process was shown to be in a validated state, as evidenced by groups, one group to be included in each of three media simula-
three successful media fills. Members of a sterility assurance tions)—and the potential contributory aseptic practices were
expert group from the wider company assisted during the investi- revised and operators retrained before conducting three success-
gation. The plant ensured that the necessary remediations ful media simulations to revalidate the process.

m a rch /a p r il 2 0 2 2 55
 TECHNICAL ASEPTIC

Table 2: Typical media fill regulatory observations.

No. Area Observations

Failure to:
• Carry out adequate growth promotion testing of media batches
• Use correct incubation conditions or duration
• Qualify all manufacturing personnel by participating in APS, and subsequently exceeding the maximum number of persons the room is qualified for
• Fill and incubate sufficient vials in the APS
1 APS design
• Simulate the lyophilization process cycle adequately
• Justify the difference between growth media makeup and pharmaceutical solution makeup
• Perform smoke studies of interventions to evaluate the effects on unidirectional (laminar) airflow
• Include representative process interventions by operators in the filling machine LAF cabinet, RABS or isolator, in the APS runs

Failure to:
• Reconcile and incubate all integral media-filled vials
2 Operational • Perform media fills after major facility shutdowns that include significant activities that may compromise cleanroom control
• Specify procedures that all personnel authorized to enter the aseptic processing rooms during manufacturing should participate in a media fill at least
once a year

Failure to:
• Determine the root cause in the investigation of APS batches exceeding the acceptance criteria for contaminated units
• Identify contaminating microorganisms to species in contaminated media units
3 Root cause analysis • Properly investigate alert or action limit exceedances in environmental monitoring, or identify contaminating microorganisms to species (such that
they can be related to microorganisms found in contaminated APS vials)
• Conduct thorough investigation on the cause of contaminated APS prior to repeating APS runs

Poor aseptic technique and practices:

• Failure to sanitize gloved hands after touching nonsterile surfaces
4 Personnel • Rapid movements in critical areas where the product is exposed to the environment
• Removing a jammed stopper by reaching over exposed sterile stoppers in the stopper bowl
• Inadequate training of media vial inspectors to examine media-filled units following incubation

Potential GMP Discrepancies During Media maintenance; and cleaning and sterilization processes. Attention
Simulations to such considerations ensures a robust and successful APS pro-
Given the enhanced frequency of regulatory inspections in com- gram.
panies where aseptic manufacturing is used and the growth of
monoclonal antibody and other biological products requiring
aseptic fi lling, there are many examples of GMP failures and APS
issues. Some typical examples that have appeared in warning let-
ters and summaries by regulators are provided in Table 2. References
1. Denyer, S. P., N. Hodges, S. P. Gorman and B. Gilmore, eds. Hugo and Russell’s Pharmaceutical
CONCLUSION Microbiology 8th Ed. Chapter 21, “Sterilisation Procedures and Sterility Assurance. 13.5
Sampling.” Wiley-Blackwell: 2011.
APS with microbial growth media is an integral part of an aseptic
manufacturing operation. The design of the APS must take into 2. China Food and Drug Administration. “Good Manufacturing Practice (2010 revision).”
Published March 2011.
consideration various operating parameters to avert a worst-case
3. China Food and Drug Administration. “Good Manufacturing Practice (2010 revision). Annex 1
scenario for the media fill challenge. Such parameters can be Sterile Medicinal Products, Article 11.” Published March 2011.
determined by risk assessment, and typically include the container-
4. China Food and Drug Administration. “Good Manufacturing Practice (2010 revision). Annex 11
closure configuration, batch size, operating conditions, and inter- Qualification and Validation.” Published March 2011.
ventions. The risks involved with individual interventions need to 5. European Commission. “EudraLex, Volume 4: EU Guidelines for GMPs for Medicinal Products
be identified, assessed, and mitigated to minimize contamination for Human and Veterinary Use. Part IV EU Guidelines for Good Manufacturing Practice (GMP)
Specific to Advanced Therapy Medicinal Products.” Published November 2017. https://ec.europa.
risk. Equally important is a team of highly trained and competent
eu/health/sites/health/files/files/eudralex/vol-4/2017_11_22_guidelines_gmp_for_atmps.pdf
operators that have knowledge of microbiology and aseptic tech-
6. European Commission. “EudraLex, Volume 4: EU Guidelines for GMPs for Medicinal Products
nique and practices; a sound and effective cleaning and disinfec- for Human and Veterinary Use. Annex 1: Manufacture of Sterile Medicinal Products.” Published
tion program for cleanrooms; regular equipment cleaning and 2008. https://ec.europa.eu/health/system/files/2016-11/2008_11_25_gmp-an1_en_0.pdf

56 P h a r m a ce u t ic a l E ngine e r ing