Sterility Assurance Strategies in Aseptic Processing for Trials

This report discusses sterility assurance in aseptic manufacturing, particularly focusing on small batches produced for clinical trials using manual aseptic processing under a laminar flow hood. It emphasizes the critical factors to be considered by sterility and quality assurance personnel before releasing a lot, including the use of laminar flow hoods and biosafety cabinets, the implementation of a robust environmental monitoring plan, and the establishment of a rigorous cleaning and gowning program. The report also highlights the importance of minimizing personnel in clean areas, providing comprehensive training, validating sterilization processes, and integrating sterile membrane filters in the filling line. Furthermore, it underscores the need for a facility maintenance program to ensure the suitability of the air handling system and the regulation of incoming components to maintain a pyrogen-free environment. The ultimate goal, irrespective of batch size or production process, is to ensure the sterility of the product throughout the entire production run.

Introduction
In the given scenario since small batches are required for clinical trial, it would not be feasible
produce them on a traditional manufacturing line. The small lots will be produced under a
laminar flow hood by manual aseptic processing. The sterility assurance of the products must be
considered carefully even when the materials are prepared through hand. However, maintaining a
suitable environment to produce aseptic drug products irrespective of the lot size is a hideous
task. The sterility and Quality assurance personnel who have the responsibility of the release of
the lot must consider several factors prior to releasing it. These factors are enumerated below:
1. It is recommended to use a laminar flow hood to undertake the operations of manual
aseptic processes. EU guidelines require a downward airflow velocity of 0.36 to 0.45 m/s.
the system should be formed to resist the pressure fluctuations within the GMP filling
room from impacting the processing space of the laminar flow hood. Laminar flow hoods
are fundamentally utilized to safeguard the product, so that their use is restricted to
products with least risk to the personnel. The flow path of the air supply from either a
horizontal in which the air is flown back to front, or a vertical in which air is flown top to
bottom, laminar flow hood exposes the personnel and the environment to the product.
The airflow from a laminar flow hood restrict its utilization if segregation of product will
be needed (Agalloco, 2017).
2. Biosafety cabinets can also be used as they offer the operators, environmental, and
product safeguarding when suitable practices and activities are followed (Fay, 2018).
With increase of Biosafety cabinets classification increases, the extent of containment
and safeguard for the personnel, product, and environment increases. Sterility assurance
of the manual aseptic processes, closed bio-safety cabinets (Class III) or modular VHP
isolators can be considered which are more sophisticated equipment substitutes. These
equipment ensure operator segregation, as the procedure will be done inside a sealed
chamber that is accessible via glove ports.
3. An established environmental monitoring plan is required (WHO, 2012). For small scale,
the plan will have information on the quality of the aseptic processing environment which
will include all the additional areas like the equipment processing area, gowning areas,

floors, ceilings, walls, and surfaces of the equipment surfaces. All the surface equipment
that contact with the product components and the product itself will be included. Suitable
monitoring sites will be ascertained by conducting a detailed risk evaluation which will
be evaluated timely and amended to include the most recent operating conditions. In
addition there will be protocol for testing final product and suitable number of samples
required to ensure sterility of the product. The samples required for testing will be taken
before, during, and after the production run and the number taken will reflect the lot size.
Small-scale parenteral lots can be manufactured successfully appropriate for patients
using several of the same processes and controls required for large-scale parenteral
production. As irrespective of the batch size or the production process, the eventual aim
is sterility assurance of the product through the complete production run. To do so, the
data gathered to ascertain the appropriateness of the processes will be used and any size
lot can be manufactured as per the regulatory authorisations (Schniepp, 2017).
4. The manufacturer company should have a vigorous plan placed to assist the sterility of
the lot irrespective of the batch size. An appropriate cleaning program which defines the
suitable utilization of disinfectants before, during, and after processing must be placed. It
can be done by having a gowning qualification program for manpower who are involved
in manufacturing of the product. This program must outline the way in which operators
are initially qualified, monitoring process for microbiological excursion during gowning,
during production, and on completing the production processes. It must also outline the
way of periodic operator’s requalification (FDA, 2015).
5. The least number of personnel needed must be present in clean areas, it is especially
relevant in the given case scenario as it includes aseptic processing. All the examinations
and controls will be done outside the clean areas to the degree that is possible. Every
personnel involved in the manufacturing process including cleaning and maintenance
personnel must get timely training in fields suitable to the accurate production of sterile
products (European Commission, 2017). The training must involve hygiene factors and
fundamental aspects of microbiology. If any worker who is not trained like building
contractors come in the area special care must be taken regarding their instruction and
monitoring. Workers who have participated in the processing of materials or of cultures

of microbes apart from those used in current manufacturing procedure must not enter
sterile-product areas until strict and well-defined entry process is followed. It is essential
that high-level protocols of personal hygiene and sanitation are maintained. People
responsible for production of sterile preparations must be taught to report the condition
which may lead to shedding of aberrant numbers or kinds of contaminants. It is also
desirable that frequent health checks for those conditions are performed. A dedicated
skilled personnel must be appointed to identify the personnel who could be introducing
undue microbial hazard and he also must make decision about what actions must be taken
on such conditions. It should be ensured that no watches, outfit and jewelry are worn in
clean areas (WHO, 2011).
6. A written procedure must be placed and followed for changing and washing. The
procedure must be planned in a way that it minimizes the contamination in clean area
clothing or transport of contaminants to the clean areas. The kind and quality of clothing
should be suitable for the procedure and the grade of the manufacturing line. Cloth must
be worn in a way that the product is protected from contamination. Since different grades
have different requirement for clothing, it must be followed accordingly. For Grade D,
hair and, if required, beard must be covered. In addition, an overall safeguarding suit and
suitable shoes must be worn. Suitable interventions will also be performed to prevent any
contamination entering from outside the working clean region. For Grade C, hair and if
required beard and moustache must be covered. Single or double piece trouser overall,
piled up at the wrists along with high neck and suitable shoes must be worn. It will be
ensured that no fibres or particulate matter should be shed virtually. For Grade A/B,
headgear must cover the hair totally and, where relevant, beard and moustache; it should
be tucked into the neck of the suit; a face mask will be worn so that shedding of droplets
can be avoided. Suitable sterilised, non-powdered gloves of plastic or rubber will be
worn. The legs of the trouser will be slipped inside the footwear and the garment sleeves
will be tucked into the gloves. The safeguard clothing must also retain the particles which
were shed by the body. Moreover, the outdoor clothing will not be carried into the
changing area directed towards Grade B and C rooms. Workers will change masks and
gloves for each working session at the least. Clean area clothing will be cleaned and

managed in a way that it does not collect extra contaminants which will be shed later.
Again, written procedures will be followed for these processes. Unsuitable management
of clothing will spoil the fibres and can enhance the risk of particle shedding.
7. Every sterilisation processes must be validated. Specific attention must be provided if the
adopted sterilization method is not outlined in the recent edition of the European
Pharmacopoeia. Caution is also required when if the method is used for a product which
is not a simple aqueous or oily solution. Heat sterilization should be preferred, if possible.
Whatever the method of sterilization is chosen, it must be in line with the marketing and
manufacturing approvals. The sterilisation process must be checked for its suitability for
the product and its effectiveness in accomplishing the preferred sterilizing conditions. In
every part of all kinds of load which has to be processed, they should be exhibited by
physical instruments and by biological indicators if suitable. Validation of the process
must be confirmed at planned time periods, minimum every year, and whenever
important alterations are done in the equipment. All the results must be documented
properly (Patel & Chotai, 2011). To make the sterilization process effective, entire
material must be treated as per the requirements and the process design must make sure
that this is accomplished. Loading patterns must be validated and established for every
sterilization activity. As mentioned earlier, biological indicators can also be regarded as
an added method for evaluating the sterilization. They must be saved and utilized as per
manufacturer’s guidelines, and their quality checked through positive controls. If
biological indicators are used for small lots, robust precautions must be taken to prevent
transfer of microorganism contamination from them. A clear way of differentiating
products must be established which separate sterilized from non-sterilized. Each basket,
tray or other carrier of products or components should be clearly labelled with the
material name, its batch number and an indication of whether or not it has been sterilised.
Indicators like autoclave tape can be utilized if suitable, to specify if a lot is passed
through a sterilisation procedure or not. But they fail to provide consistent indication that
the batch is, actually, sterile. Records of sterilization must be available for all the
sterilization runs. They should be accepted as an aspect of the lot release process.

8. High intrinsic value products which will be used in trials can be tested and manufactured
in small unit batches of less than 20 containers, can be integrated with the filling
activities. A sterile membrane filter will be integrated in the filling line and the containers
are filled and several containers are sampled. Containers which are sampled are spun to
make sure that the product touches the complete inner surface of the container. After that
aseptic procedures are used and the containers are unbolted and the contents of the
containers are evacuated into the filling line reservoir. After that the product will be
refilled into new containers and the in-line membrane filter is detached and sterility test is
done by distributing it into two nearly same quantity and testing the two portions in two
different mediums. The number of containers utilized for filling the medium must be
adequate to facilitate a valid evaluation. In the given case scenario, since small batches
are required, then the number of containers filling the medium should minimally equal
the size of the product lot. Zero growth should be the target and since between 200 and
500 units are manufactured, it is standardized that no contaminated units must be
discovered.
9. A facility maintenance program will also be there which will challenge the suitability of
the air handling system. A timely maintenance program will be established for high-
efficiency particulate air (HEPA) air filtering system and frequent smoke studies will be
performed to ensure the suitability of the airflow for aseptic processing. It will give the
assurance regarding the appropriateness of the airflow system control and functioning.
10. Apart from the above mentioned considerations, a plan will also be required to regulate
incoming components. All the components that will be utilized in the production of
aseptic processing will need be delivered pyrogen free prior to their introduction into the
production area. The small-scale lots will not be given the advantage of continuous
activities related to large-scale lot production, therefore, it the process of introducing the
materials and components manually into the production area will be considered.
Conclusion
Therefore, it can be concluded that various sterility assurance approach are standard irrespective
of the lot size. However, in the given case scenario since laminar flow hood by manual aseptic

processing is used, certain procedures are necessary, like personnel hygiene, gowning and
changing room etc. As irrespective of the batch size or the production process, the eventual aim
is sterility assurance of the product through the complete production run.
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