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AUDIT OF

STERILE PRODUCT

MANUFACTURING UNIT

Sandesh Deshpande

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Quality Assurance Department,

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Serum Institute of India Pvt Ltd.,

STERILE PRODUCTION

• Manufacture of Sterile Preparations:

The manufacture of sterile products is subject to special requirements in order to minimize risks of microbial, particulate and Pyrogen (endotoxin) contamination.

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Principle

• Facility, equipment, and process design should be optimized, qualified, and validated according to the Good Manufacturing Practices (GMP)
• The use of appropriate technologies (e.g. Restricted Access Barriers Systems RABS), isolators and robotic systems.
• Appropriate written procedures(SOP, Policy etc. ), designed to prevent microbiological contamination of drug products purporting to be sterile, shall be established and followed.
• Personnel should have adequate qualification & experience training and attitude with a specific focus on principles involved in the protection of the sterile product during manufacturing.

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Point to be considered

• Points to be considered during sterile product audit.
• Premise
• Disinfection
• Equipment
• Utilities.
• Personnel
• Aseptic preparation & processing
• Aseptic process simulation (APS)

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PREMISES

• Manufacture of sterile products there are four grades of cleanroom (Clean area air classification ).
• Grade A:
• The critical zone for high risk operations.
• Sterile starting materials and components. Preparation of solutions (if not to be sterile filtered later) Handling and filling of aseptically prepared products, Transfer of partially closed containers, before complete stoppering.
• Grade B area:
• For aseptic preparation and filling, this is the background cleanroom for the Grade A zone.
• Grade C & D area
• These are cleanrooms used for carrying out less critical stages in the manufacture of aseptically filled sterile products but can be used for the preparation /filling of terminally sterilized products.(background for isolator)
• Clean Non Classified (CNC ) area.

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PREMISES

• In cleanrooms, all exposed surfaces should be smooth in order to minimize the generation of particles.
• Sinks and drains are prohibited in Grade A zone and Grade B area.
• The transfer of materials, equipment, and components into an aseptic processing area should be carried out via a unidirectional process.
• Pass-through hatches should be designed to protect the higher grade environment, for example by effective flushing with an active filtered air supply

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PREMISES

• Personnel airlocks: Areas of increasing cleanliness used for entry of personnel (e.g. from Grade D to Grade C to Grade B).
• Material airlocks: used for materials and equipment transfer.
• Appropriate Biosafety cabinet /isolators shall be used for handling of live organism.
• Dedicated Autoclave for sterilisation & decontamination (Inactivation).

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� DISINFECTION

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• Disinfectants shall be selected based on their chemistry, compatibility and ability to reduce the bioburden.

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• Each disinfectant shall be qualified for its efficacy at the concentration and contact time recommended by the manufacturer.

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• This validated concentration and contact time shall be used for routine cleaning.

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• Disinfectants used in Grade A & B zone should be sterilised.

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• Disinfectants shall be rotated among the class (chemistry) of them on defined frequency, which may be include sporicidal disinfectant based on its effect on product.

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• Fumigation or vapour disinfection (e.g. Vapour-phased Hydrogen Peroxide) of cleanrooms and associated surfaces may be useful for reducing microbial contamination in inaccessible places.

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EQUIPMENT

• Direct and indirect contact parts should be sterilized. Direct contact parts are those that the product passes through, such as filling needles or pumps.
• Indirect product contact parts are equipment parts that come into contact with sterilized critical items and components.
• All the equipment such as sterilizers, air handling systems, and water systems should be subjected to qualification, monitoring &planned maintenance
• Particle counters, including sampling tubing, should be qualified. The tubing length should be no greater than 1 meter with a minimum number of bends.
• Portable particle counters with a short length of sample tubing should be used for classification purposes.
• Isokinetic sampling heads should be used in unidirectional airflow systems and should be positioned as close as possible to sample air representative of the critical location.

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UTILITIES

• Utilities should be designed, installed, qualified, operated, maintained and monitored in a manner to ensure that the utility functions as expected.
• Results for critical parameters and critical quality attributes of high-risk utilities should be subject to regular trend analysis to ensure that system capabilities remain appropriate.
• Gases that come in contact with the product should be tested for chemical, particulate & microbial quality.
• Gases used in aseptic processes should be filtered through sterilizing grade filter

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Personnel

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• Only the minimum number of personnel required should be present in cleanrooms.
• All personnel performing cleaning, maintenance, monitoring in cleanrooms should receive regular training, gowning qualification and assessment in disciplines relevant to the correct manufacture of sterile products.
• The personnel working in a Grade A zone and Grade B areas should be trained for gowning and aseptic practices.
• Clothing should be chosen to prevent shedding due to operators moving excessively (when cold) or sweating (when hot).

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Aseptic preparation & processing

• Products that cannot be filtered,

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All product and component contact equipment should be sterilized prior to use.

All raw materials should be sterilized and aseptically added or subsequently sterilized by filtration.

Bulk solutions should be sterilized by a validated process, e.g., heat, sterilization or sterile filtration.

All materials added to the sterile bulk product should be sterilized prior to addition.

Use of single-use equipment.

Use of close connectors

Aseptic preparation & processing

• The duration of each aspect of aseptic preparation should be limited and validated
• Holding time of cleaned equipment.
• Sterilized equipment hold time.
• Hold time study of the product prior to filling.
• Aseptic processing time
• Holding time of decontaminated environment such as isolator
• Filling time

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ASEPTIC PROCESS

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VALIDATION

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QA & REGULATORY PERSPECTIVE

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Demonstrate the capability of the aseptic process to produce sterile drug products.

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Evaluate the proficiency of aseptic processing personnel.

Comply with current Good Manufacturing Practice requirements.

PROCESS SIMULATION /� ASEPTIC PROCESS VALIDATION

• Process Simulation means that microbiological nutrient media will be filled into a container closure system (Prefilled Syringes/ Ampoules and Vials) instead of the product under simulation of aseptic standard procedure

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objective of aseptic simulation

• The industry and practice and standards reflects philosophy that the media fill program is designed.

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• To ensure that media fill are conducted under “worst case” conditions.

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• The worst case conditions provide challenge to aseptic condition.

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• The objective of worst case is not to result in failure but to provide the high degree of assurance.

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STUDY DESIGN

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NUMBER & FREQUENCY OF RUN

• Three initial runs including routine and non- routine intervention for initial validation.
• Semi-annual re-conformation of studies.

Additional simulation may be required based on:

• Modifications to the equipment.
• Modification to equipment or facilities that potentially affects

the air quality or airflow in the aseptic environment.

• Major changes in the number of production personnel or initiation of second (or third) shift production.
• Any major shutdown.
• Any product sterility test failure.
• Continued critical area environmental monitoring results above the alert/action levels.

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• Each media fill run should evaluate a single line speed and speed chosen should be justified.
• Using the slowest fill speed for the largest container (maximum opening).
• Using the highest fill speed for the small container (handling difficulty/ more interventions).

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LINE SPEED

CONTAINER SIZE

• Initial qualification carry out three consecutive separate successful media fill runs using container & closure configuration of worst case.
• Largest size container with a wide opening and subsequently three media fill runs using the smallest pack size, which covers entire range of container closure system.
• Routinely media fill run shall contain any one container size, which is altered every 6 months to cover minimum and maximum size containers.

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SET-UP

• The filling line set-up usually entails manual assembly of equipment.
• Equipment set-up activities may require more manipulation of critical surfaces than subsequent filling operation.
• The process simulation test to be designed to detect the potential contamination from set-up activities.

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CONTAINER CLOSURE SYSTEM

• In case of unique operating challenges

(e.g., tipping, jams) and causes increased

interventions, it is recommended a separate

process simulation be performed with that

particular configuration.

• Clear containers of identical configuration may be substituted for opaque or amber containers to aid in the detection of contamination.

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FILL VOLUME

• Regardless of the actual fill volume, the process simulation should include a fill weight / volume adjustment using methods identical to those employed during production.
• There are two general criteria.
• First, there must be enough medium in the container to contact all the container-closure seal surfaces when the container is inverted and swirled.
• Second, there must be in the container to allow for the detection of microbial growth.
• The volume of headspace should be considered in the growth promoting capability of the media to support aerobic microorganisms.

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DURATION

• Routine aseptic interventions such as initial setting, adjustment of fill volume, shift change, manual maintenance of equipment should be included in the process simulation study design.

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• For high speed filling it is usually appropriate to fill additional units in order to accommodate normal aseptic manipulations and intervention.

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• Duration of the media fill should be not less than that of duration of filling of regular batch.

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INTERVENTION

An aseptic manipulation or activity performed by personnel that occurs with in critical area.

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• Inherent interventions:

Inherent interventions are normal and planned activities that occur during aseptic process.(e.g. equipment set-up, weight adjustment,EM monitoring).

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• Corrective interventions:

Corrective intervention are performed to correct or adjust an aseptic process during its execution.it is not part of planned aseptic process.

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���� �����������������INCUBATION �

• Process simulation units should be incubated for a

minimum of 14 days.

• Inverted unit prior to each incubation.
• The temperature chosen should be based upon its ability to recover microorganisms normally found environmentally or in the product bioburden.

Two temperature are used for incubation of media fill

units, the units should be incubated at least 7 days at

each temperature (starting with lower temperature:

20-25°C and 30-35°C for at least 07 days each)

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��Media Selection and Growth Promotion test

• The most common medium for process simulation is Soybean-Casein Digest Broth (SCDM).
• Sterilised by either steam or filtration.
• Pre-incubated SCDM media shall be used for simulation.
• The media selected should be demonstrated to promote the growth of gram positive and negative bacteria and yeast and mold after 14 days of incubation.
• Environmental monitoring and sterility test isolates can be added to growth promotion challenge.
• Growth promotion unit should be inoculated with a

<100 cfu challenge.

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INERT GAS

• The sterility of the inert gas system is confirmed through filter validation and integrity testing, not by means of the process simulation.
• Where sterile inert gases are used during normal production, process simulation test should be substituted with sterile air.
• The use of an inert gas with Soybean-Casein Digest Medium may inhibit growth.

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PERSONNEL CONSIDERATION

• Each person who work in aseptic filling area should be participate in successful media fill trials on periodic basis.

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• However participation only is not enough, operator must performed same tasks they would perform in the execution of normal fill.

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WORST CASE

The use of worst case situation is intended to provide the greater challenge to process, system or equipment being validated.

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However, worst case condition should not be deliberately used in regular production, because they are validated.

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Example of Worst Case

• Use of materials, components and closures which have remained in clean area for extended periods.

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• On particular line, filling the smallest units at faster speed (more intervention) and the largest unit at slowest operating speed. (more exposure).

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• Maximum number of personnel present in clean room.

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�INSPECTION OF MEDIA FILL

• The unit should be examined visually for evidence of growth.
• Each media fill units should be visually checked for contamination by personnel with appropriate training for inspection of microbial contaminating unit.
• Damaged containers should not be consider for evaluation of aseptic processing capability of process.
• If the container is non integral, the source of breach should be investigated and corrective action taken.

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INTERPRETATION OF RESULTS AND ACCEPTANCE CRITERIA

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Despite the numbers of unit filled during process simulation test or number of positive allowed, the ultimate goal for number of positives in any process simulation test should be zero.

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ACCEPTANCE CRITERIA

• When filling fewer than 5000 units,

No contaminated units should be detected.

One contaminated unit should result in an investigation,

including consideration of a repeat media fill.

• When filling 5,000 to 10,000 units:
• One (1) contaminated unit should result in an investigation, including consideration of a repeat media fill.
• Two (2) contaminated units are considered cause for revalidation, following investigation.

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ACCEPTANCE CRITERIA

• When filling more than 10,000 units:
• One (1) contaminated unit should result in an investigation;
• Two (2) contaminated units are considered cause for revalidation, following investigation

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Incubation period and evaluating results

• If any vial is found contaminated during the scheduled inspection, identify the vial/tray and record the details in the BMR for media fill at respective place.
• All suspected vials identified during the examination period, should be brought to the immediate attention of QA.
• At the end of incubation period, failure investigation to be done followed by these vials should be examined in laboratory, for contamination characterization / identification.

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INVESTIGATION APPROCH

Review of Mfg. Data : -

• Media preparation and Filtration.
• Review of Batch manufacturing record
• Clean Room Entry and Exit
• Disinfectant preparation and sanitization Record
• Sterilization of Garment, Product contact material and primary packaging material
• Filter integrity Record.

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INVESTIGATION APPROCH

• Environment Monitoring Record –
• Viable, Non-viable (online/offline), Temperature, Relative Humidity and pressure differential
• Critical Alarm Record
• Video recording activity.
• Deviation and Change control record

Personnel Record:

• Gowning qualification
• Training Records.
• Handling of intervention.

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INVESTIGATION APPROCH

Quality Control Review.

• Viable and non-viable environmental monitoring data.
• Positive container(s) identification up to genus and species level
• Comparison of identified isolate with the database of recently identified organisms
• OOL record if any
• Sterility Test failure history if any
• Media sample results – GPT, Sterility and Container closure integrity .
• Empty vial/ampoule, Rubber stopper sterility Test reports
• EM monitoring plates release data.

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INVESTIGATION APPROCH

Engineering Aspect :-

• Maintenance activity before/ during media fill activity (if any)
• Engineering tools /material transfer during media fill (if any)
• HVAC qualification.
• WFI, Pure steam and Compressed Air Qualification.
• Lyophilizer sterilization record and any maintenance activity
• Last Qualification DHS. Autoclave validation, Lyophilizer ( If applicable).Via washing, filling machine and sealing machine.

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INVESTIGATION APPROCH

Sampling Plan:-

• Primary packing material (vials and Rubber stopper sample) for sterility test.
• Priming sample for incubation (VDMG) and GPT.
• Left Over samples for incubation (VDMG).
• Seal integrity test of incubated Vials.
• Post GPT of incubated Vials.

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INVESTIGATION REPORT

• Based upon the outcome Of the investigation. the cause Of the failure is either assignable or not assignable.
• If the cause is assignable, corrective action needs to be taken and documented.
• Process simulations required to demonstrate that the process is operating Within the expected parameters.
• Where assignable cause cannot be determined, and considering previous process simulation results. multiple consecutive successful simulation may be required to reconfirm the aseptic process control.

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INVESTIGATION REPORT

The investigation report should contain:

• •A summary Of the occurrence.
• •A list Of the systems investigated.
• •A conclusion as to root cause(s) and supporting scientific data (rational /Justification)
• •Potential effect on previous batches produced.
• •Corrective action(s) taken and supporting data
• •Outcome Of additional proces simulations, if performed
• •Appropriate signatures report should be signed by Production QA, and QC .

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REFERENCE(S)

• Guidance for Industry - Sterile Drug Products Produced by Aseptic Processing - Current Good Manufacturing Practice, USFDA, Sept. 2004.

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• PIC/S validation of aseptic process.

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• WHO TRS 961 Annex - 6.

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• PDA Technical Report No. 22.

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• D & C ACT Schedule M

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• EU Annex-1

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Thank You

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