A3P 16th Annual conference

November 2012

Assuring sterility A CMOs approach

Introduction
Patheon Inc. is a leading global provider of contract development and manufacturing services to the global pharmaceutical industry. We provide products and services to approximately 300 of the world's leading pharmaceutical and biotechnology companies. Our services range from preclinical development through commercial manufacturing of a full array of solid and sterile dosage forms, including softgels. As the site Microbiologist at Patheon UK, (based in Swindon) I am responsible for the Microbiology and sterility assurance functions at two sites, serving a total of 10 sterile filling lines and 3 non sterile filling lines for both commercial and pre-clinical development batches.

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Assuring sterility Introduction

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Assuring sterility A CMOs approach

Our goal in the production process is to assure the absence of microorganisms from every container. However, what available means do we have to demonstrate such an absolute negative? Sterility testing - is severely limited statistically (20 containers tested per batch)

Media fills (process simulations) - are point in time assessments of the capabilities of an aseptic process and even then only support a maximum contamination rate, not a level of sterility assurance.
Environmental monitoring - can only recover a small percentage of the microorganisms present. (contact plates recover an average of 29% of S. epidermidis from a stainless steel surface)

Absence of growth does not mean the absence of microorganisms!

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Assuring sterility A CMOs approach

Drug product

Sterilisation

Sterile drug product

Container

Sterilisation

Sterile Container
Aseptic processing

Sterile final product

Closure

Sterilisation

Sterile Closure

Excipient

Sterilisation

Sterile Excipient

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Assuring sterility A CMOs approach

The essence of aseptic processing is the assembly of previously sterilised materials into a sealed container using procedures, controls and equipment that preserves their most critical attribute - sterility

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Assuring sterility A CMOs approach

Six blind men and an elephant!

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Assuring sterility A CMOs approach

The six pillars of a robust aseptic process:

Facilities design HVAC validation Material / component transfer Process simulation (media fills) Personnel training & monitoring Environmental monitoring
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Assuring sterility A CMOs approach

Like the elephant, the aseptic process must be considered holistically

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Assuring sterility Facility and design

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Assuring sterility Facility and design

In terms of the general design of cleanrooms, it is imperative that:

They are built of an airtight structure

As a minimum it should be an ISO 5 / Grade A environment The internal surfaces should be smooth and suitable for cleaning The internal surface finish should be sufficiently resilient to resist chipping or powdering The surfaces should be resistant to the cleaning agent used. The designs for manned cleanrooms have undergone constant revisions with many of these designed to increase the separation between the operator and the aseptic portions of the process. The culmination of this progression is the closed isolator which affords the highest degree of physical separation from the operator.
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The Aseptic Processing Spectrum

Reducing probability of units being contaminated

Open Gowned Operator Access

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Increasing confidence in sterility assurance Confidential

Assuring sterility Facility and design

RABS = Restricted Access Barrier System Process improvements like RABS are aimed at separating people from the process. If successful the risk posed by people contaminating the area will be greatly reduced

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Assuring sterility Facility and design

Patheon RABS approach
HEPA HEPA HEPA HEPA
Grade A

HEPA

HEPA

Open operation
Doors can be opened but only for predefined / risk assessed activities

Grade B traditional cleanroom

Fixed Barriers Glove ports

Filling Line

Overspill of Grade A Air below the critical zone of operation

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Assuring sterility HVAC

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Assuring sterility HVAC

The HVAC (Heating, Ventilation and Air Conditioning) system controls the room parameters including temperature and humidity. This is important not only for operator comfort but also for minimising microbial contamination.
High temperatures can lead to excessive perspiration which can reduce the efficiency of the cleanroom suit and result in the increase of shedding of micro-organisms Certain mould species cannot grow in areas of low humidity, therefore keeping the room humidity low will ensure that contamination will be kept to a minimum.

Guidance would suggest that the temperature should be maintained at 18oC 2oC and humidity at 45% 15
We use a building management system to control these parameters constantly to show that the room is in control

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Assuring sterility - HVAC

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Assuring sterility Material and Component transfer

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Assuring sterility Material and Component transfer

Materials and components should wherever possible enter your cleanroom as sterile, in order to achieve this they can go via a hierarchy of controls which aims to eradicate the microbial contamination.
Autoclaving usually at 121oC for a minimum of 15 minutes (validated by demonstration of a 106 log reduction of a biological indicator) De-pyrogenation (validated by demonstration of a reduction of endotoxin level) Gamma irradiation usually 25kGy (validated by dose mapping) Ethylene Oxide - (validated by demonstration of a 106 log reduction of a biological indicator)

Whatever method you use you must ensure that all sterilisation and sanitisation processes are properly validated Recognise that premium equipment, package components and materials should offer enhanced reliability over lesser quality items. Aim to reduce the assembly time required for all sterilised materials through pre-fabrication or use of disposable assemblies.

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Assuring sterility Material and Component transfer

How the material has been sterilised will determine how it enters the cleanroom: In the ideal situation some components will be steam sterilized directly into the Grade B cleanroom (the problem here will be how to get it to the Grade A environment (we use Grade A mobile LAF carts)). Many others arrive pre-sterilized (irradiation or EtO) and enter the via a series of airlocks.

Patheon procedure requests that components arrive sealed inside 3 PE bags. This allows us to operate an effective decontamination process that controls the risks posed by the outer surfaces of these items.
Eg. Post sterilization the components usually arrive inside a cardboard box; cardboard can get damp during transport and can allow proliferation of moulds.

Moulds are a risk to our processes.

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Assuring sterility Material and Component transfer

Process for transfer of pre-sterilised components:
Remove Cardboard Box Vacuum clean if dust present Disinfect Bag Remove Outer Bag AND Disinfect Inner Bag Remove Outer Bag Or Disinfect Outer Bag

Support Area

Grade C

Grade B

Grade A (eg RABS)

Contamination on outer surface

No contamination

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Assuring sterility Material and Component transfer Component Chute

HEPA

HEPA

Component chute pivots to open & close Positioned relatively high to prevent operator leaning in

Components opened in outer A and poured down the chute. Chute closes immediately

FILLING LINE

Overspill of Air

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Assuring sterility Media fill simulations

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Assuring sterility Media fill simulations

The main contamination risk evaluation tool capable of including all the different aspects and variables of the aseptic process is a process simulation (or media fill). The design of a media fill simulation should consider the following factors: Media selection Frequency and number of runs Size and duration of runs Line speed, container size, closure type and fill volume Process/line configuration set up Number of persons, activities and interventions (including line clearance)

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Assuring sterility Media fill simulations

Choosing the media

The media fill should mimic as closely as possible your production of product.
If you have a powder filling process you should use a sterile placebo (we use PEG, polyethylene glycol) As most clean room contaminants are strict or facultative aerobes any step in the process where oxygen content is kept low could cause a potential growth inhibition and could mask contamination routes. (ie by the use of Nitrogen). If your process does use Nitrogen then this should be substituted during the media fill with compressed air. If this is not possible then evidence that growth of typical process contaminants is not affected by this step should be considered. If on the other hand your manufacturing process is a totally anaerobic process then you should consider the use of an anaerobic media such as Alternative Fluid Thioglycollate medium
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Assuring sterility Media fill simulations

Frequency and number of runs At least three consecutive separate successful runs are necessary for: Initial line qualification

Revalidation following major changes to the equipment and/or process

Whenever there are doubts about the ability of the aseptic process to exclude contamination (extended shutdowns, trends in environmental monitoring, sterility test failures etc) For routine re-qualification, one run per line with a semi-annual frequency (or twice a year, as reported in the Annex 1) is recommended.

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Assuring sterility Media fill simulations

Size and duration of runs

Ideally, the most accurate simulation would be to mimic exactly the commercial process in terms of batch size and total duration. In practice, provided that they address all the potential risks of contamination occurring during commercial production a different model can be used.
5,000 to 10,000 units is a generally acceptable starting point for run size (FDA guidance), however, for commercial products with batch sizes much bigger than 10,000 units it could be difficult to justify that a simulation run smaller than 10,000 units is considered representative of the actual process. The run size must be big enough to allow all of the manipulations and interventions occurring during the commercial process to be captured.

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Assuring sterility Media fill simulations

Size and duration of runs

From a practical point of view the problem is how to combine a duration equal or longer than the longest commercial process with a media fill run size that is much smaller than the product batch size.
At Patheon we perform the process simulation running the line intermittently taking care to include line set up, all the planned interventions and maximum operator levels. Interventions Interventions are generally divided into two main categories: Intrinsic to the process (normal interventions) Extrinsic to the process (non-routine interventions)

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Assuring sterility Media fill simulations

Routine Interventions These include all the interventions that are necessary for running the process as it is designed. Line set up Aseptic connection(s) Container/Closure feeding Product sampling Weight checks

Environmental monitoring

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Assuring sterility Media fill simulations

Non routine interventions These include all the other interventions that are not strictly necessary for running the process, but that can occur due to expected or unexpected problems during the operation. Removal of jammed containers/closures Removal of broken containers/product residues Replacement of a dosing device Adjustment to the line set-up (guides, star wheels etc) Breakdowns (sensors, switches, belts etc)

The perfect intervention is one that doesnt exist

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Assuring sterility Media fill simulations

Interventions into the RABS
A standard list of pre assessed interventions is published for each RABS

Each new intervention must be risk assessed prior to being performed.

A. Amount of microbial contamination on source; 0-2 (eg a sterilized tool or a persons hand) B. Risk of dispersion, 0-2 (vigorous movements required. Working above the product or product handling?) C. Proximity to product, 0-2 (distant from the product or close by?) D. Effectiveness of control measures, 0-2 (localised clean, discard of exposed product & components) Defines controls required e.g. Terminate the batch Localised clean

AxBxC

Risk posed by intervention

AxBxCxD

Residual risk after control measures taken

H, M, L or N

Defines the EM Required e.g. Take fingerdabs Take extra sterility samples

Duration defined
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Assuring sterility Media fill simulations

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Assuring sterility Facility and design Example of an intervention

Codes are located on each door of the RABS These are the only routine interventions that the operator can perform.

The number and types of interventions are compared during batch release to those that have been performed during a Media Fill

H 301 M 302 M 303 L 304 N 305

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Colour indicates Risk and indicates how to perform the intervention Letters indicates the residual risk after the corrective actions are taken and specifies what EM is required Numbers are linked to a written description of the intervention within the SOP and the batch record

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Assuring sterility Facility and design Intervention Matrix

Colour = Risk
Do not record Sanitise Hands Use glove ports Use tools

Blue

Record in BMR Sanitise Hands Use glove ports or hatch Use tools

Green

Record in BMR Sanitise Hands Open door or use hatch Use tools

Orange

Record in BMR Sanitise Hands Open door or use hatch Use hands

Red

Also how it is done

Code = Risk remaining after the intervention Code specifies the EM and samples required Maximum allowed time is specified

N = negligible
No EM or Samples

L = Low
No EM or Samples

M = Medium
Finger-dabs

Finger-dabs, sterility test samples

H = High

Duration Exceeded No action required

Duration Exceeded BNF

Duration Exceeded BNF

Duration Exceeded BNF + Risk Assessment

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Media fill
Intervention

checklist

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Assuring sterility Training of personnel

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Assuring sterility Training of personnel

The biggest source of contamination in a cleanroom is people, and levels of contamination will be highest when their training is poor. Gowns, masks, gloves and goggles can reduce the incidence of contamination but the risk remains. The closer the people get to the product and the more they interact with the line and the product the more likely that contamination will be transferred. It is therefore essential that adequate training is conducted before an individual is allowed to enter the cleanroom to perform aseptic manipulations.

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Assuring sterility Training of personnel

As a minimum the training of the operator should include education in: GMP Personal hygiene Gowning and comportment training

Basic Microbiology
Knowledge of how a cleanroom functions The impact their behaviours can have on the product

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Assuring sterility Training of personnel

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Assuring sterility Training of personnel

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Assuring sterility Training of personnel

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Assuring sterility Training of personnel

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Assuring sterility Training of personnel

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Assuring sterility Training of personnel

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Assuring sterility Environmental monitoring

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Introduction
The Patheon site at Swindon has ten Grade A (Class 100) manufacturing lines. Environmental monitoring is performed at defined frequencies and locations using Settle plates, Active air samples and contact plates. The locations for the EM monitoring has been defined based on risk assessment of the operation in conjunction with smoke visualisation footage and trend data.

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Environmental monitoring - Frequency

Grade
Settle plates Contact plates Active air sample Routine Finger Dab Intervention Finger Dab Sleeve monitoring

A
Continuous monitoring End of each batch/campaign Every shift After every exit After every high and medium risk intervention At the end of each entry a high risk intervention is performed After every exit End of each batch/campaign

B
Each operational shift Daily Each operational shift After every exit N/A

N/A
After every exit N/A

Gown Monitor Swabs

(An operational shift is defined as an 8 hour period)

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Environmental monitoring Choosing the location

EM locations are primarily selected based on an assessment of the process with respect to risk from microbial contamination using the following equation: Risk from microbial contamination (Risk rating) = A x B x C x D Where: A = Microbial contamination on, or in, a source.

B = Ease of dispersion and transfer of contamination.

C = Proximity and ease of contaminating source from the critical area. D = Effectiveness of contamination control method.

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Environmental monitoring Choosing the location

A committee should be set up to carry out the risk assessment and comprise the following people: Risk management expert Microbiologist Quality Assurance manager Facilities manager Production manager

The risk scores should be assigned to hazards by an aggregation of opinion from the committee individuals and you should all establish a method of scoring to suit your facility.
We use the scoring system 0 2 with

0 = nil, 0.5 = very low, 1.0 = low, 1.5 = medium and 2.0 = high
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Environmental monitoring Choosing the locations

Environmental monitoring is increased or decreased based on the risk assessment score. ie Red ratings would require environmental monitoring to be performed during that activity or intervention

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Environmental monitoring Choosing the locations

The number of points to be sampled in a given area should be related to risk, therefore the sampling points should be concentrated in the critical areas, and as the risk of contamination in the surrounding areas becomes less (ie corridors and other support areas) then there should be fewer sampling points. Transfer areas may be the exception and require more sampling because of the higher risk associated with the materials and personnel that moving to the critical area.

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Environmental monitoring Choosing the locations

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Environmental monitoring Choosing the locations

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Assuring sterility Conclusion

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Assuring sterility Conclusion

Like the elephant, the aseptic process must be considered holistically

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Thank you for listening!

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References
Risk Management of Contamination (RMC) During Manufacturing Operations In Cleanrooms Technical Monograph No. 14 by the Pharmaceutical and Healthcare sciences society (PHSS) and the Scottish society for contamination control

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