Commission Implementing Regulation (EU) 2025/2091 of 17 October 2025 laying down good manufacturing practice for veterinary medicinal products in accordance with Regulation (EU) 2019/6 of the European Parliament and of the Council

VIII.4.7.Procedures shall be in place for the assessment and interpretation of appropriate actions (where required) in light of the results obtained from the sampling. Supporting data for the recovery efficiency of the sampling methods chosen shall be available. Action limits for viable particle contamination are shown in Table 6.

Grade	Air sample CFU/m3	Settle plates (diam. 90 mm) CFU/4 hours (1)	Contact plates (diam. 55mm), CFU/plate (2)	Glove print, Including 5 fingers on both hands CFU/glove
A	No growth (3)
B	10	5	5	5
C	100	50	25	—
D	200	100	50	—
(1) — Settle plates shall be exposed in grade A and B areas for the duration of operations (including equipment set-up) and changed as required after a maximum of 4 hours (exposure time shall be based on validation including recovery studies and it shall not have any negative effect on the suitability of the media used). — For grade C and D areas, exposure time (with a maximum of 4 hours) and frequency shall be based on quality risk management principles. — Individual settle plates may be exposed for less than 4 hours. (2) Contact plate limits apply to equipment, room and gown surfaces within the grade A and grade B areas. Routine gown monitoring is not normally required for grade C and D areas, depending on their function. (3) For grade A, any growth shall trigger an investigation. Note 1:  The types of monitoring methods listed in the table above are examples and other methods may be used provided they are capable of providing information across the entire critical process where the product may be contaminated (e.g. aseptic line set-up, aseptic processing, filling and lyophilizer loading). Note 2:  Limits are applied using CFU throughout the document. If different or new technologies are used that present results in a manner different from CFU, the manufacturer shall scientifically justify the limits applied and where possible correlate them to CFU.

VIII.4.8.Microorganisms detected in the grade A and grade B areas shall be identified to the species level and the potential impact of such microorganisms on product quality (for each batch implicated) and overall state of control shall be evaluated. The identification of microorganisms detected in grade C and D areas shall be considered, as appropriate, as part of the contamination control strategy (for example where action limits or alert levels are exceeded) or following the isolation of organisms that may indicate a loss of control or deterioration in cleanliness, or the isolation of organisms that may be difficult to control such as spore-forming microorganisms and moulds and at a sufficient frequency to maintain a current understanding of the typical flora of these areas.

VIII.5.   Aseptic process simulation (also known as media fill) (³⁶)

VIII.5.1.The periodic verification of the effectiveness of the controls in place for aseptic processing shall include an aseptic process simulation using a sterile nutrient media and/or a surrogate in place of the product. The selection of the nutrient media and/or the surrogate shall be made based on the ability of the media and/or the surrogate to imitate physical product characteristics posing a risk to the product sterility during the aseptic process. Where processing stages may indirectly impact the viability of any introduced microbial contamination, (e.g. aseptically produced semi-solids, powders, solid materials, microspheres, liposomes and other formulations where product is cooled, heated or lyophilised), alternative procedures that represent the operations as closely as possible shall be developed. Where surrogate materials, such as buffers, are used in parts of the aseptic process simulation, the surrogate material shall not inhibit the growth of any potential contamination.

The aseptic process simulation is not the primary means to validate the aseptic process or aspects of the aseptic process. The effectiveness of the aseptic process shall be determined through process design and process controls, training, and evaluation of monitoring data.

VIII.5.2.The aseptic process simulation shall imitate as closely as possible the routine aseptic manufacturing process and include all the critical manufacturing steps, specifically:

(a) the aseptic process simulation shall assess all aseptic operations performed subsequent to the sterilisation and decontamination cycles of materials utilised in the process to the point where the container is sealed;

(b) for non-filterable formulations, any additional aseptic steps shall be assessed;

(c) where aseptic manufacturing is performed under an inert atmosphere, the inert gas shall be substituted with air in the process simulation unless anaerobic simulation is intended;

(d) processes requiring the addition of sterile powders shall use an acceptable surrogate material in the same containers as those used in the process under evaluation;

(e) separate simulations of individual unit operations (e.g. processes involving drying, blending, milling and subdivision of a sterile powder) shall be avoided. Any use of individual simulations shall be supported by a documented justification and ensure that the sum total of the individual simulations continues to fully cover the whole process;

(f) the process simulation procedure for lyophilised products shall represent the entire aseptic processing chain including filling, transport, loading, a representative duration of the chamber dwell, unloading and sealing under specified, documented and justified conditions representing worst case operating parameters;

(g) the lyophilisation process simulation shall mimic all aspects of the process, except those that may affect the viability or recovery of contaminants. For instance, boiling-over or actual freezing of the solution shall be avoided. Factors to consider in determining aseptic process simulation design include, where applicable: — the use of air to break vacuum instead of nitrogen or other process gases; — replicating the maximum interval between sterilisation of the lyophilizer and its use; — replicating the maximum period of time between filtration and lyophilisation; — quantitative aspects of worst-case situations, e.g. loading the largest number of trays, replicating the longest duration of loading where the chamber is open to the environment.

VIII.5.3.The aseptic process simulation shall take into account the aseptic manipulations and interventions known to occur during normal production as well as worst-case situations, as well as the following:

(a) inherent and corrective interventions representative of the routine process shall be performed in a manner and frequency similar to that during the routine aseptic process;

(b) the inclusion and frequency of interventions in the aseptic process simulation shall be based on the assessed risks posed to the product sterility.

VIII.5.4.The aseptic process simulation shall not be used to justify practices that pose unnecessary contamination risks.

VIII.5.5.The following elements are relevant for the development of the aseptic process simulation plan:

(a) identification of worst-case conditions covering the relevant variables, such as container size and line speed, and their impact on the process. The outcome of the assessment shall justify the variables selected;

(b) identification of the representative sizes of container/closure combinations to be used for validation. A bracketing or matrix approach may be considered for validation of the same container/closure configuration for different products where the process equivalence is scientifically justified;

(c) identification of the maximum permitted holding times for the product and for the equipment exposed during the aseptic process;

(d) identification of the volume filled per container, which shall be sufficient to ensure that the media contacts all the equipment and component surfaces that may directly contaminate the product. The volume used shall also provide sufficient headspace to support potential microbial growth and ensure that turbidity can be detected during inspection;

(e) substitution of any inert gas used in the routine aseptic manufacturing process by air, unless anaerobic simulation is intended. In this case, the inclusion of occasional anaerobic simulations as part of the overall validation strategy shall be considered as appropriate;

(f) the selected nutrient media shall be capable of growing a designated group of reference microorganisms as described by the relevant pharmacopeia and suitably representative local isolates (³⁷);

(g) the method of detection of microbial contamination shall be scientifically justified to ensure that contamination is reliably detected;

(h) the process simulation shall be of sufficient duration to challenge the process, the operators that perform interventions, shift changes and the suitability of the processing environment;

(i) where the manufacturer operates different or extended shifts, the aseptic process simulation shall be designed to capture factors specific to those shifts that may pose a risk to product sterility, for example the maximum duration for which an operator may be present in the cleanroom;

(j) simulating normal aseptic manufacturing interruptions where the process is idle (e.g. shift changeovers, recharging dispensing vessels, introduction of additional equipment);

(k) ensuring that the environmental monitoring is conducted as required for routine production and throughout the entire duration of the process simulation;

(l) where campaign manufacturing occurs, such as in the use of barrier technologies or manufacture of sterile active substances, consideration shall be given to designing and performing the process simulation so that it simulates the risks associated with both the beginning and the end of the campaign and demonstrating that the campaign duration does not pose any risk;

(m) the performance of ‘end of production or campaign aseptic process simulation’ may be used as additional assurance; however, such approach cannot replace routine aseptic process simulation.

VIII.5.6.For sterile active substances, the batch size shall be large enough to represent the routine operation, simulate operation at the worst case, and cover all surfaces that may come into contact with the sterile product. In addition, all the simulated materials (surrogates or growth medium) shall be subject to microbial evaluation. The simulation materials shall be sufficient to ensure the robustness of the evaluation of the process being simulated and shall not compromise the recovery of microorganisms.

VIII.5.7.Aseptic process simulation shall be performed as part of the initial validation, with at least three consecutive satisfactory simulation tests that cover all working shifts that the aseptic process may occur in. In addition, an aseptic process simulation is also mandatory after any significant modification to operational practices, facilities, services or equipment that may have an impact on the sterility assurance of the product (e.g. modification to the HVAC system, the equipment, changes to process, the number of shifts or number of personnel, or after a major facility shut down). Moreover, an aseptic process simulation (periodic revalidation) shall usually be repeated twice a year (approximately every six months) for each aseptic process, each filling line and each shift. Each operator shall participate in at least one successful aseptic process simulation annually. Consideration shall be given to performing an aseptic process simulation after the last batch prior to shut down, before long periods of inactivity or before the decommissioning (i.e. definitively removing from the manufacturing process) or the relocation of a line.

VIII.5.8.Where manual operation occurs (e.g. aseptic compounding or filling), each type of container, container closure and equipment train shall be initially validated with each operator participating in at least 3 consecutive successful aseptic process simulation and revalidated with one aseptic process simulation approximately every 6 months for each operator. The aseptic process simulation batch size shall mimic the one used in the routine aseptic manufacturing process.

VIII.5.9.The number of units processed (filled) for aseptic process simulation shall be sufficient to effectively simulate all the activities that are representative of the aseptic manufacturing process. Justification for the number of units to be filled shall be addressed as part of the contamination control strategy. Typically, a minimum of 5 000 to 10 000 units shall be filled. For small batches (e.g. those under 5 000 units), the number of containers for aseptic process simulation shall at least equal the size of the production batch.

VIII.5.10.Filled aseptic process simulation units shall be agitated, swirled or inverted before incubation to ensure contact of the media with all interior surfaces in the container. All integral units from the aseptic process simulation shall be incubated and evaluated, including units with defects without a critical impact on the integrity of the container (e.g. those with cosmetic defects) or those which have gone through non-destructive in-process control checks.

If units are discarded during the process simulation and not incubated, those shall be comparable to the units discarded during a routine fill, and only if the standard operating procedures applicable to production specify that units must be removed under the same circumstances (i.e. type of intervention, line location, specific number of units removed). Under no circumstances may more units be removed during a media fill intervention than during a production run. Examples may include those that shall be discarded during routine production after the set-up process or following a specific type of intervention.

VIII.5.11.Where the manufacturing process includes materials that are in contact with the product surface but are then discarded (e.g. product flushes), the discarded material shall be simulated with nutrient media and be incubated as part of the aseptic process simulation, unless it can be clearly demonstrated that this waste process does not have an impact on the sterility of the product.

VIII.5.12.Filled aseptic process simulation units shall be incubated in a clear container to ensure visual detection of microbial growth. Where the product container is not clear (e.g. amber glass, opaque plastic), clear containers of identical configuration may be substituted to aid in the detection of contamination. When a clear container of identical configuration cannot be substituted, a suitable method for the detection of microbial growth shall be developed and validated. Microorganisms isolated from contaminated units shall be identified to the species level where possible, to assist in the determination of the likely source of the contaminant.

VIII.5.13.Filled aseptic process simulation units shall be incubated without unnecessary delay to achieve the best possible recovery of potential contamination. The selection of the incubation conditions and duration shall be scientifically justified and validated to provide an appropriate level of sensitivity for the detection of microbial contamination.

VIII.5.14.Upon completion of incubation, filled aseptic process simulation units shall be inspected by personnel who have been appropriately trained and qualified for the detection of microbiological contamination. Such inspection shall be conducted under conditions that facilitate the identification of any microbial contamination. In addition, samples of the filled units shall undergo a positive control by inoculation with a suitable range of reference organisms and suitably representative local isolates.

VIII.5.15.The target is zero growth. Any contaminated unit shall be considered as a failed aseptic process simulation and the following actions shall be taken:

(a) investigation to determine the most probable root cause(s);

(b) determination and implementation of appropriate corrective measures;

(c) a sufficient number of successful, consecutive aseptic process simulations (normally a minimum of 3) shall be conducted in order to demonstrate that the process has been returned to a state of control;

(d) a prompt review of all appropriate records relating to aseptic production since the last successful aseptic process simulation shall be made. The outcome of the review shall include a risk assessment of the potential breaches in batches manufactured since the last successful aseptic process simulation. In addition, all other batches not released to the market shall be included in the scope of the investigation. Any decision regarding their release status shall take into account the investigation outcome;

(e) all products that have been manufactured on a line subsequent to a process simulation failure shall be quarantined until a successful resolution of the process simulation failure has occurred;

(f) where the root cause investigation indicates that the failure was related to operator activity, actions to limit the involved operator’s activities, until retrained and requalified, shall be taken;

(g) production shall resume only after completion of successful revalidation.

VIII.5.16.All aseptic process simulation runs shall be fully documented and include a reconciliation of the units processed (e.g. units filled, incubated and not incubated). Justification for the filled and the non-incubated units shall be included in the documentation. All interventions performed during the aseptic process simulation shall be recorded, including the start and the end time of each intervention and the involved person(s). All microbial monitoring data as well as other testing data shall be recorded in the aseptic process simulation batch record.

VIII.5.17.An aseptic process simulation run shall only be aborted under circumstances in which written procedures require commercial lots to be equally handled. An investigation shall be performed and documented in such cases.

VIII.5.18.The validation of the aseptic process shall be repeated when the specific aseptic process has not been in operation for an extended period of time or when there is a change to the process, the equipment, the procedures or the environment that has the potential to affect the aseptic process, or when new product containers or container closure combinations are added.

SECTION IX

QUALITY CONTROL

IX.1.There shall be personnel available with appropriate training and experience in microbiology, sterility assurance and knowledge of the processes to support the design of the manufacturing activities, environmental monitoring regime and any investigation assessing the impact of microbiologically linked events to the safety of the sterile product.

IX.2.Specifications for raw materials, components and products shall include requirements for microbial, particulate and endotoxin/pyrogen limits when considered necessary having regard to the monitoring data and the overall contamination control strategy.

IX.3.The bioburden assay shall be performed on each batch for both aseptically filled product and terminally sterilised products and the results shall be taken into consideration as part of the final batch review. Limits for bioburden immediately before the final sterilising grade filter or the terminal sterilisation process shall be set having regard to the efficiency of the method to be used. Samples shall be taken to be representative of the worst-case scenario (e.g. at the end of hold time). Where overkill sterilisation (³⁸) parameters are set for terminally sterilised products, bioburden shall be monitored at suitable scheduled intervals.

IX.4.For products authorised for parametric release, a supporting pre-sterilisation bioburden monitoring programme for the filled product prior to initiating the sterilisation cycle shall be developed and the bioburden assay shall be performed for each batch. The sampling locations of filled units before sterilisation shall be based on a worst-case scenario and be representative of the batch. Any organisms found during the bioburden testing shall be identified and their impact on the effectiveness of the sterilising process determined. Where appropriate, the level of endotoxin/pyrogen shall also be monitored.

IX.5.The sterility test applied to the finished product shall be validated for the product concerned. This test is only the last in a series of critical control measures by which sterility is assured and it may not be used to ensure sterility of a product that does not meet the relevant design, procedural or validation parameters.

IX.6.The sterility test shall be performed under aseptic conditions. In addition, samples taken for sterility testing shall be representative of the whole batch but shall in particular include samples taken from parts of the batch considered to be most at risk of contamination, for example:

— for products which have been filled aseptically, samples shall include containers filled at the beginning and end of the batch. The taking of additional samples shall be considered based on risk (e.g. after critical interventions);

— for products that have been heat sterilised in their final containers, samples taken shall be representative of the worst case locations (e.g. the potentially coolest or slowest to heat part of each load);

— for products that have been lyophilised, samples shall be taken from different lyophilisation loads.

Note:Where the manufacturing process results in sub-batches (e.g. for terminally sterilised products), samples from each sub-batch shall be taken and a sterility test performed for each sub-batch. Where appropriate, consideration shall be given to performing separate testing for other finished product tests.

IX.7.When it is not possible to have the sterility test result prior to release because the shelf life of the product is too short, additional process controls and monitoring and/or alternative test methods implemented to mitigate the identified risks shall be scientifically justified and documented.

IX.8.Any process (e.g. vaporised hydrogen peroxide, ultra violet) used to decontaminate the external surfaces of the sterility samples prior to testing shall not negatively impact the sensitivity of the test method or the reliability of the sample.

IX.9.Media used for product testing shall be quality control tested according to the Pharmacopeia before use. Media used for environmental monitoring and aseptic process simulation shall be tested for growth promotion before use, using a scientifically justified and designated group of reference microorganisms and including suitably representative local isolates. Media quality control testing shall usually be performed by the end user. Reliance on outsourced testing or supplier testing of media shall be justified and transportation and shipping conditions be duly considered.

IX.10.Environmental monitoring data and trend data generated for classified areas shall be reviewed as part of the product batch certification/release. A written procedure shall be available describing the actions to be taken when data from environmental monitoring are found out of trend or exceeding the established limits. For products with a short shelf life, where the environmental data for the time of manufacture is not available, a review of the most recent available data is required. In addition, the use of rapid/alternative methods may be considered.

IX.11.Where rapid and automated microbial methods are used in manufacturing, those methods shall be validated for the product(s) or processes concerned.

ANNEX II

I. SCOPE

I.1.The additional requirements set out in this Annex shall apply to the manufacture, control and testing of biological and immunological veterinary medicinal products, with the exception of inactivated immunological veterinary medicinal products which are manufactured from pathogens and antigens obtained from an animal or animals in an epidemiological unit and used for the treatment of that animal or those animals in the same epidemiological unit or for the treatment of an animal or animals in a unit having a confirmed epidemiological link.

Throughout the Annex, reference to ‘biological veterinary medicinal products’ or ‘biologicals’ is to be understood as encompassing also immunologicals.

I.2.Antibiotics are generally not considered biologicals. Nevertheless, manufacturers are advised to follow the requirements set out in this Annex with regard to manufacturing procedures described in this document that are used in the manufacture of such veterinary medicinal products.

I.3.Table 1 illustrates the manufacturing activities which are generally within the scope of this Annex.

Type and source of material	Example product	Manufacturing steps covered by this Annex shown in grey GMP requirements shall increase from the earlier steps (e.g. collection) to the final manufacturing steps (formulation, filling). For earlier stages of manufacturing, GMP principles shall at least be adhered to.
Human source	Urine derived enzymes, hormones	Collection of material (1)	Mixing, and/or initial processing	Isolation and purification	Formulation, filling
Animal or plant sources (not genetically modified/edited)	Heparins, insulin, enzymes, proteins, allergen extract, immunosera	Collection of plant or animal material (2)	Cutting, mixing, and/or initial processing	Isolation and purification	Formulation, filling
Virus or bacteria /fermentation/cell culture, etc.	Viral or bacterial vaccines; enzymes, proteins	Establishment & maintenance of MCB, WCB, MSL, WSL (3)	Cell culture and/or fermentation	Inactivation when applicable, isolation and purification	Formulation, filling
Biotechnology- fermentation/cell culture	Recombinant products, MAb, allergens, vaccines	Establishment & maintenance of MCB and WCB, MSL, WSL (4)	Cell culture and/or fermentation	Isolation, purification, modification	Formulation, filling
Animal sources, (genetically modified/edited)	Recombinant proteins	Master and working (genetically modified/edited) bank	Collection, cutting, mixing, and/or initial processing	Isolation, purification, modification	Formulation, filling
Plant sources: (genetically modified/edited)	Recombinant proteins, vaccines, allergen	Master and working (genetically modified/edited) bank	Growing, harvesting (5)	Initial extraction, isolation, purification, modification	Formulation, filling
(1) GMP principles shall be adhered to. (2) See Section IV for the extent to which GMP principles apply. (3) See Section VI for the extent to which GMP applies. (4) See Section VI for the extent to which GMP applies. Maintenance of working cell bank is expected to take place in a GMP environment. (5) The standards of good agricultural and collection practice for starting materials of herbal origin (GACP) is applicable.

I.4.In cases where there is a continuous process from the sourcing/isolation of the active substance from a biological source to the manufacturing of the finished product (e.g. veterinary medicinal products that consists of cells, viral-based vaccines, phages), the requirements of this Regulation shall apply to the entire manufacturing process.

II.   PERSONNEL

II.1.Personnel (including those concerned with cleaning and maintenance) employed in areas where biological products are manufactured and tested shall receive initial and periodic retraining specific to the products manufactured and their respective tasks, including measures to protect the product, personnel and the environment, as well as – where appropriate – training on microbiology.

II.2.Personnel shall be protected against a possible infection with the biological agents used in the manufacture. In the case of biological agents known to cause disease in humans, adequate measures shall be taken to prevent infection of personnel working with the agent or with experimental animals. Where appropriate, relevant vaccination and health monitoring shall be offered having regard to the specific characteristics of the manufactured product (e.g. BCG vaccine (³⁹), rabies, brucella, leptospira, tuberculin products) and the tasks of the staff.

II.3.When a heath condition that may have a negative impact on the quality of the product is declared by the relevant personnel or otherwise becomes apparent, access to the production or control area shall be barred.

II.4.Where required to minimise the opportunity for cross-contamination, restrictions on the movement of all personnel (including quality control, maintenance and cleaning staff) shall be applied on the basis of quality risk management principles (⁴⁰). In general, personnel shall not pass from areas where exposure to live micro-organisms, genetically modified/genome edited organisms, toxins or animals occurs to areas where other products, or different organisms are handled. If such passage is unavoidable, appropriate contamination control measures commensurate to the risks shall be implemented.

II.5.Adequate measures shall be implemented to prevent biological agents being taken outside the manufacturing plant by personnel acting as a carrier. Dependent on the type of biological agent, such measures may include a complete change of clothes and compulsory showering before leaving the production area.

III.   PREMISES AND EQUIPMENT

III.1.   Premises

III.1.1.Premises shall be designed in such a way as to control the risks to the product and the risks to the environment. This may be achieved by the use of contained, clean, or controlled areas. In particular:

(a) Live biological agents that are highly pathogenic shall be handled in contained areas (including quality control operations, research and diagnostic services, etc). The level of containment shall be adapted to the pathogenicity of the agent.

(b) Without prejudice to the containment measures that may be required for certain biological organisms as provided for under (a), production of biological agents may generally take place in controlled areas provided it is carried out in totally enclosed and sterilised equipment and all relevant connections are made in accordance with the requirements set out in Annex I.

(c) Inactivated biological agents shall be handled in clean areas. Clean areas shall also be used when handling non-infected cells isolated from multicellular organisms and, where appropriate, filtration-sterilised media.

(d) Open circuit operations involving products or components not subsequently sterilised shall be carried in accordance with the requirements set out in Annex I, where relevant.

(e) With the exception of blending and subsequent filling operations (or when closed systems are used), only one biological agent shall be handled at a given time within an area.

(f) Production operations such as cell maintenance, media preparation, virus culture, etc. which are likely to cause contamination shall be performed in separate areas, unless appropriate organisational and technical measures may be put in place to prevent contamination.

(g) Production areas where biological agents particularly resistant to disinfection (e.g. spore-forming bacteria) are handled shall be separated and dedicated to that particular purpose until the biological agents have been inactivated, unless appropriate organisational and technical measures may be put in place to prevent contamination.

III.1.2.As part of the contamination control strategy, the degree of environmental control of particulate and microbial contamination of the production premises shall be adapted to the active substance, intermediate or finished product and the production step, bearing in mind the potential level of contamination of the starting materials and the risks to the product. Where relevant in accordance with quality risk management principles, the environmental monitoring programme shall be supplemented by the inclusion of methods to detect the presence of specific microorganisms (i.e. host organism, yeast, moulds, anaerobes, etc.).

III.1.3.Manufacturing and storage facilities, processes and environmental classifications shall be designed to prevent the extraneous contamination of products. Where processes are not closed and there is therefore exposure of the product to the immediate room environment (e.g. during additions of supplements, media, buffers, gases) adequate control measures shall be put in place, including engineering and environmental controls. Relevant aspects addressed in Annex I shall be applied, including aspects related to the required environmental grades and associated controls.

III.1.4.Air handling units shall be designed, constructed and maintained to minimise the risk of cross-contamination between different manufacturing areas, to provide relevant containment where applicable, and may need to be specific for an area. Consideration, based on quality risk management principles, shall be given to the use of single pass air systems.

III.1.5.Positive pressure areas shall be used to process sterile products but negative pressure in specific areas at the point of exposure of pathogens is acceptable for containment reasons. Where negative pressure areas or safety cabinets are used for aseptic processing of materials with particular risks (e.g. pathogens) they shall be surrounded by a positive pressure clean zone of appropriate grade. These pressure cascades shall be clearly defined and continuously monitored with appropriate alarm settings.

III.1.6.Air vent filters shall be hydrophobic and validated for their scheduled life span with integrity testing at appropriate intervals based on appropriate quality risk management principles.

III.1.7.Where necessary to address the contamination risks, equipment passes and changing rooms shall have an interlock mechanism or other appropriate system to prevent the opening of more than one door at a time. Changing rooms shall be supplied with air filtered to the same standard as that for the work area and equipped with air extraction facilities to produce an adequate air circulation independent of that of the work area. Equipment passes shall normally be ventilated in the same way, but unventilated passes, or those equipped with supply air only, may be acceptable.

III.1.8.Drainage systems shall be designed so that effluents can be effectively neutralised or decontaminated to minimise the risk of cross-contamination. Local regulation shall be followed to minimise the risk of contamination of the external environment according to the risk associated with the biohazardous nature of waste materials.

III.1.9.Containment premises shall be designed so as to ensure that they can be easily disinfected and shall have the following characteristics:

(a) Absence of direct venting to the outside.

(b) Ventilation with air at negative pressure: Air shall be extracted through HEPA filters and not be re circulated except to the same area, and provided further HEPA filtration is used (normally this condition would be met by routing the recirculated air through the normal supply HEPAs for that area). Recycling of air between areas is acceptable only if it passes through two exhaust HEPAs, the first of which is continuously monitored for integrity, and there are adequate measures for safe venting of exhaust air should this filter fail. Where negative pressure areas are used for containment purposes of aseptic processing of materials with particular risks, they shall be combined with a positive pressure clean zone of appropriate grade (pressure bubble or sink airlock). These pressure cascades shall be clearly defined and continuously monitored with appropriate alarm settings.

(c) By way of derogation from (b), air from manufacturing areas used for the handling of exotic organisms (⁴¹) shall be vented through two sets of HEPA filters in series, and air from production areas shall not be re-circulated.

(d) A system shall be put in place for the collection and disinfection of liquid effluents including the contaminated condensate from sterilisers, biogenerators, etc. In addition, solid wastes, including animal carcasses, shall be disinfected, sterilised or incinerated as appropriate. Contaminated filters shall be removed using a safe method.

(e) Changing rooms shall be designed and used as air locks and equipped with washing and showering facilities if appropriate. Air pressure differentials shall be such that there is no flow of air between the work area and the external environment or a risk of contamination of the outer clothing worn outside the area.

(f) An airlock system is in place for the passage of equipment, which is constructed so that there is no flow of contaminated air between the work area and the external environment or a risk of contamination of the equipment within the lock. The airlock shall be of a suitable size to enable the effective surface decontamination of materials being passed through it. Consideration shall be given to having a timing device on the door interlock to allow sufficient time for the decontamination process to be effective.

(g) Where appropriate, a barrier double-door autoclave shall be used for the secure removal of waste materials and introduction of sterile items.

III.1.10.The measures and procedures implemented for containment to address operator and environmental safety shall not conflict with those required to ensure product quality.

III.1.11.Manufacture of biologicals in a multi-product facility is acceptable where appropriate measures are implemented to prevent cross-contamination, such as:

— measures to prevent cross-contamination to non-related areas or equipment (e.g. use of single use components and engineering measures such as closed systems);

— validated cleaning and decontamination procedures before the subsequent manufacture of other products, covering also heating, ventilation and air conditioning (HVAC) system;

— environmental monitoring in adjacent areas – during manufacture and after completion of cleaning and decontamination – specific for the micro-organism being manufactured. Risks arising from the use of certain monitoring equipment (e.g. airborne particle monitoring) in areas handling live and/or spore-forming organisms shall be duly considered;

— controls on the movement/removal of products, equipment, ancillary equipment (e.g. for calibration and validation) and disposable items to prevent contamination of other areas, other products or different product stages (e.g. prevent contamination of inactivated or toxoided products with non-inactivated products);

— campaign-based manufacturing.

Where production involves the manufacture of multiple small batches from different starting materials, factors such as the health status of donors and the risk of total loss of product shall be taken into account when considering the acceptance of concurrent working.

III.1.12.For finishing operations (formulation, filling and packaging), the assessment whether dedicated facilities are required shall also take into consideration the specific characteristics of the concerned biological product and the characteristics of other products, including any non-biological products, handled in the same facility.

Other control measures for finishing operations may include the need for specific addition sequences, mixing speeds, time and temperature controls, limits on exposure to light, as well as containment and cleaning procedures in the event of spillages.

III.2.   Equipment

III.2.1.Equipment used during handling of live organisms and cells, including equipment used for sampling, shall be suitable to prevent contamination during processing, facilitate decontamination and sterilisation (where applicable), and to avoid any mix-up between different organisms or products.

Particular attention shall be paid to control measures to avoid cross-contamination from parts of equipment that are not fixed, such as pipes, valves and filters (e.g. appropriate identification as to their function).

III.2.2.Equipment used for the storage of biological agents or products shall be suitable and used in such a manner as to prevent any possible mix-up. All stored items shall be clearly and unambiguously labelled and in leak-proof containers.

III.2.3.Where relevant, equipment shall be fitted with recording and/or alarm systems (e.g. equipment requiring temperature control). To avoid breakdowns, a system of preventive maintenance, together with trend analysis of recorded data, shall be implemented.

III.2.4.Close equipment used for primary containment shall be designed and periodically tested to ensure the prevention of escape of biological agents into the immediate working environment. Inlets and outlets for gases shall be protected so as to achieve adequate containment, e.g. by the use of sterilising hydrophobic filters. The introduction or removal of material shall take place using a sterilisable closed system, or possibly in an appropriate laminar air flow.

III.2.5.Where necessary, equipment shall be properly sterilised by a validated method. Equipment used for purification, separation or concentration shall be sterilised or disinfected at least between each use for different products. The effect of the sterilisation methods on the effectiveness and validity of the equipment shall be studied in order to determine the life span of the equipment.

III.2.6.The use of ‘clean in place’ (⁴²) and ‘steam in place’ (⁴³) (‘sterilisation in place’) systems shall be used where appropriate. Valves on fermentation vessels shall be completely steam sterilisable.

III.2.7.The loading of freeze dryers requires an appropriate clean/contained area. Unloading freeze dryers contaminates the immediate environment. Therefore, for single-ended freeze dryers, the clean room shall be decontaminated before a further manufacturing batch is introduced into the area, unless that batch involves the same organism(s). Double door freeze dryers shall be sterilised after each cycle unless opened in a clean area. Sterilisation of freeze dryers shall be done in accordance with Annex I. In case of campaign working, they shall at least be sterilised after each campaign.

IV.   ANIMALS

The use of animals or animal materials in the manufacture of veterinary medicinal products is subject to specific additional requirements:

(a) Biological substances and products shall comply with the latest version of the Note for guidance on minimising the risk of transmitting animal spongiform encephalopathy agents via human and veterinary medicinal products.

(b) Specifications for materials of animal origin shall consider aspects such as age, weight and health status of the animals, as appropriate, and in accordance with the terms of the marketing authorisation.

(c) Where applicable, pharmacopeia requirements shall be complied with, including the need for specific tests at defined stages.

(d) A health programme shall be established to monitor adventitious agents that are of concern (zoonotic diseases, diseases of source animals). Specialist advice shall be obtained for the establishment of such programme. In particular, reports from trustworthy sources on national disease prevalence shall be considered when performing the assessment of risk and mitigation factors. Such sources include the World Organisation for Animal Health (WOAH) (⁴⁴), as well as information on health monitoring and control programme(s) at national and local level.

(e) Instances of ill-health occurring in the source/donor animals shall be investigated with respect to their suitability and the suitability of in-contact animals for continued use (in manufacture, as sources of starting and raw materials, in quality control and safety testing). Decisions shall be documented. A look-back procedure shall be in place to inform the decision-making process on the continued suitability of the biological active substance or medicinal product in which the animal sourced starting or raw materials have been used or incorporated. This decision-making process may include the re-testing of retained samples from previous collections from the same donor animal (where applicable) to establish the last negative donation. The withdrawal period of therapeutic agents used to treat source/donor animals shall be documented and used to determine the removal of those animals from the programme for defined periods.

(f) Particular care shall be paid to the prevention and monitoring of infections in the source/donor animals, including addressing aspects regarding the sourcing, the facilities, the husbandry, the biosecurity procedures, the testing regimes or the control of bedding and feed materials. This is of special relevance to specified pathogen free (⁴⁵) animals where PhEur monograph requirements shall be met. Housing and health monitoring shall be defined for other categories of animals (e.g. healthy flocks or herds).

(g) For products manufactured from animals that have been genetically modified or whose genome has been edited, traceability shall be maintained in the creation of such animals from the source animals.

(h) Housing for animals used in production and control of biological active substances and medicinal products shall be separated from production and control areas. Such houses shall be provided with the appropriate containment and/or clean area measures and shall be separate from other animal accommodation. It is particularly important to ensure that animal houses where animals used for quality control involving the use of pathogenic biological agents are accommodated are adequately contained.

(i) At the production site, animals, biological agents and tests carried out on them shall be duly identified to prevent any risk of confusion and to control all the identified hazards.

(j) Suitable measures shall be implemented to ensure the quality and the traceability of materials of animal origin sourced from abattoirs. In particular, the contract/supply agreement with the abattoir shall address the required measures to ensure traceability of sourced materials, as well as the implementation of adequate hygiene and other necessary control measures at the abattoir.

(k) Adequate controls (based on quality risk management principles) shall be applied over the supply chain and during the transport of animals or animal materials used in the manufacture of veterinary medicinal products, including detailed documentation to ensure traceability. Traceability shall be ensured, including the movement of material between sites of initial collection, partial and final purification(s), storage sites, hubs, consolidators and brokers. Details of such arrangements shall be recorded and any breaches recorded, investigated and actions taken.

V. STARTING AND RAW MATERIALS

V.1.The source, origin and suitability of biological starting and raw materials (e.g. cryoprotectants, feeder cells, reagents, culture media, buffers, serum, enzymes, cytokines, growth factors) shall be clearly defined in written specifications. Specifications shall include quality requirements necessary to ensure the suitability of the materials for the intended use and to minimise variability (covering relevant aspects of the production and control). Microbiological controls are particularly important. The specifications set shall be in compliance with the terms of the marketing authorisation. Starting and raw materials shall be appropriately identified through the various stages of production.

V.2.Where the results from the test(s) required to release the starting materials take a long time (e.g. sterility test), it may be permissible to process the starting materials before the results of the test(s) are available, provided that the risk of using a potentially failed material and its potential impact on other batches is understood and assessed under quality risk management principles. In such cases, the release of a finished product shall be conditional on the satisfactory results of these tests.

V.3.The risk of contamination of the starting and raw materials during their passage along the supply chain shall be assessed, with particular emphasis on TSE. Materials that come into direct contact with the manufacturing equipment or with the product (such as media used in media fill experiments and lubricants that may contact the product) shall also be taken into account.

V.4.A control strategy to protect the product and the preparation of solutions, buffers and other additions based on the principles set out in Annex I shall be implemented. The controls required for the quality of starting and raw materials and on the aseptic manufacturing process are particularly important for products in respect of which final sterilisation is not possible.

V.5.Where sterilisation of starting and raw materials is required, it shall be carried out – where possible – by heat. Where necessary, other appropriate methods may also be used for inactivation of biological materials (e.g. irradiation and filtration).

V.6.The use of antibiotics at early manufacturing stages to reduce bioburden (e.g. bioburden associated with the procurement of living tissues and cells) shall generally be avoided. The use thereof shall be duly justified. In such cases, the presence of antibiotics shall be removed from the manufacturing process at the stage specified in the marketing authorisation.

V.7.Adequate measures shall be implemented throughout the supply chain to ensure the traceability of substances of animal and human origin that are used in the manufacture of veterinary medicinal products.

Donor (human or animal) health information having an impact on the quality of the veterinary medicinal product which becomes available after procurement shall be taken into account in recall procedures.

VI.   SEED LOT AND CELL BANK SYSTEM

VI.1.In order to prevent the unwanted drift of properties that may result from repeated subcultures or multiple generations, the production of biological substances and products obtained by microbial culture, cell culture or propagation in embryos and animals shall be based on a system of master and working seed lots (⁴⁶) and/or cell banks (⁴⁷).

VI.2.The number of generations (doublings, passages) between the seed lot or cell bank, the active biological substance and the finished product shall be consistent with the specifications in the marketing authorisation.

VI.3.As part of the product lifecycle management, the establishment of seed lots and cell banks, including master and working generations, shall be performed under circumstances which are demonstrably appropriate, including a controlled environment which is adequate to protect the seed lot and the cell bank and the personnel handling it. In addition, during the establishment of the seed lot and cell bank, no other living or infectious material (e.g. virus, cell lines or cell strains) shall be handled simultaneously in the same area or by the same persons.

VI.4.For stages prior to the master seed or cell bank generation, where only the principles of GMP may be applied, documentation shall be available to support traceability, including regarding components used during the development with a potential impact on product safety (e.g. reagents of biological origin), from initial sourcing and genetic development if applicable. For vaccines, compliance with the requirements in Pharm. Eur. monograph on Vaccines for veterinary use 01/2023:0062 is required.

VI.5.Following the establishment of master and working cell banks and master and working seed lots, quarantine and release procedures shall be followed, including adequate characterisation and testing for contaminants. Their on-going suitability for use shall be further demonstrated by the consistency of the characteristics and quality of the successive batches of product. Evidence of the stability and recovery of the seeds and banks shall be documented and records shall be kept in a manner permitting trend evaluation.

VI.6.Seed lots and cell banks shall be stored and used in such a way as to minimise the risks of contamination (e.g. stored in the vapour phase of liquid nitrogen in sealed containers) or alteration. Control measures for the storage of different seeds and/or cells in the same area or equipment shall prevent mix-up and take into account the infectious nature of the materials to prevent cross contamination.

VI.7.Storage containers shall be sealed, clearly labelled and kept at an appropriate temperature. A stock inventory shall be kept. The storage temperature shall be recorded continuously and, where used, the liquid nitrogen level monitored. Deviation from set limits and corrective and preventive action taken shall be recorded.

VI.8.It is desirable to split stocks and to store the split stocks at different locations so as to minimise the risks of total loss. The controls at such locations shall provide the assurances outlined in the preceding paragraphs.

VI.9.The storage and handling conditions for stocks shall be managed according to the same procedures and parameters. Once containers are removed from the seed lot/cell bank management system, the containers shall not be returned to stock.

VII.   PRODUCTION

VII.1.Quality risk management principles shall be implemented across all the stages of the manufacture of biological veterinary medicinal products to minimise process variability and to enhance reproducibility. The effectiveness of the implemented measures shall be reassessed during product quality reviews.

VII.2.Critical operational (process) parameters and other input parameters that affect product quality shall be identified, validated, documented and be shown to be maintained within required parameters.

VII.3.Changes introduced in the manufacturing process shall comply with the requirements laid down in Article 26(3). In addition, the cumulative effects of changes introduced in the manufacturing process on the quality, safety and efficacy of the finished product shall be evaluated on periodic basis.

VII.4.Where starting materials from different donors are used, adequate controls shall be implemented to minimise the risk of cross-contamination or mix-ups.

VII.5.For biological materials that cannot be sterilised (e.g. by filtration), processing shall be conducted aseptically to minimise the introduction of contaminants. The requirements on aseptic manufacturing set forth in Annex I shall be implemented.

VII.6.The terms of the marketing authorisation or – as appropriate – Pharmacopoeia monographs, shall determine whether, and up to what stage, substances and materials used in the manufacturing of biological veterinary medicinal products can have a defined level of bioburden or need to be sterile. Appropriate controls shall be implemented to ensure that the specified limits are respected.

VII.7.Appropriate measures shall be implemented throughout all the production stages and controls to prevent or minimise the occurrence of unwanted bioburden and associated metabolites and endotoxins.

VII.8.A control strategy for the entry of articles and materials into production areas shall be implemented based on quality risk management principles. The following shall be implemented where applicable:

(a) For aseptic processes, heat stable articles and materials entering a clean/contained area (⁴⁸) shall preferably do so through a double-ended autoclave or oven. Heat labile articles and materials shall enter through an airlock with interlocked doors where they shall be subject to effective surface sanitisation procedures. The sterilisation of articles and materials elsewhere is acceptable provided that multiple wrappings are used, as appropriate to the number of stages of entry to the clean area, and enter through an airlock with the appropriate surface sanitisation precautions.

(b) Equipment, glassware, the external surfaces of product containers and other such materials shall be disinfected before being transferred from a contained area using a validated method. Only the absolute minimum of materials shall enter or leave the area.

(c) Liquid or solid wastes such as the debris after harvesting eggs, disposable culture bottles, unwanted cultures or biological agents, shall preferably be sterilised or disinfected before being transferred from a contained area. However, alternatives such as the use of sealed containers or piping may be appropriate in some cases.

(d) Where relevant/critical raw materials (such as culture media and buffers) have to be measured or weighed during the production process (e.g. due to variability concerns), small stocks of these raw materials may be kept in the production area for a specified duration based on defined criteria (e.g. for the duration of manufacture of the batch or of the campaign).

VII.9.The growth promoting properties of culture media shall be demonstrated to be suitable for its intended use. If possible, media shall be sterilised in situ.

VII.10.The addition of materials or cultures to fermenters and other vessels and sampling shall be carried out under carefully controlled conditions to prevent contamination and, in case of live micro-organism, egress. It shall be verified that vessels are correctly connected when the addition or the sampling takes place. Gases, media, acid or alkalis, defoaming agents and other materials introduced into sterile biogenerators shall be sterile where appropriate.

VII.11.Continuous monitoring of some production processes, such as e.g. fermentation, may be necessary (e.g. continuous monitoring of parameters such as temperature, pH, pO2, CO2 and the rate of feed or carbon source with respect to growth of cells) and such data shall form part of the batch record. Special consideration shall be given to the quality controls required when continuous culture is used.

VII.12.The formation of droplets and the production of foam shall be avoided or minimised as far as possible during manufacturing. Centrifugation and blending of products can lead to aerosol formation. Therefore, such activities shall be adequately contained with a view to minimise the risk of cross-contamination or, where relevant, risks to operators or the environment.

VII.13.Accidental spillages, especially of live organisms, shall be dealt with quickly and safely. Validated decontamination measures shall be available for each organism or groups of related organisms. Where different strains of single bacteria species or very similar viruses are involved, the decontamination process may be validated with one representative strain, unless there is reason to believe that they may vary significantly in their resistance to the agent(s) involved.

VII.14.Production and control materials, including paperwork, that are obviously contaminated, such as by spills or aerosols, or if a potential hazardous organism is involved, shall be adequately decontaminated, or the information be transferred out by other means.

VII.15.Precautions shall be taken to avoid contamination or confusion during incubation. Separate incubators shall be used for infected and non-infected containers and also generally for different organisms or cells. Incubators containing more than one organism or cell type are only acceptable if adequate steps are taken to seal, decontaminate the surface and segregate the containers. Culture vessels and any other container shall be carefully and clearly labelled. Specific cleaning/decontamination procedures for incubators shall be laid down.

VII.16.In cases where a virus inactivation or a removal process is performed during manufacture, measures shall be taken to avoid the risk of recontamination of treated products by non-treated products. Vessels containing inactivated products shall not be opened or sampled in areas containing live biological agents. For sterile products and for aseptic manufacturing, handling shall be done in accordance with Annex I.

VII.17.The inactivation process for live organisms shall be validated. For products that are inactivated by the addition of a reagent (e.g. micro-organisms in the course of vaccine manufacture) the process shall ensure the complete inactivation of the live organism and prevent any subsequent contamination from any equipment surface.

VII.18.Where chromatography equipment is used in campaign manufacture and in multi-product environments, a suitable control strategy (based on risk management principles) for matrices, the housings and associated equipment shall be implemented. The re-use of the same matrix at different stages of processing is discouraged. When it occurs, such re-usage shall be supported by appropriate validation data. Acceptance criteria, operating conditions, regeneration methods, life span, and sanitisation or sterilisation methods of chromatography columns shall be defined.

VII.19.Applicable requirements when irradiated equipment and materials are used are laid down in Annex VII.

VII.20.Filling shall be carried out as soon as possible following production. Containers of bulk product prior to filling shall be sealed, appropriately labelled and stored under specified conditions of temperature.

VII.21.When there is a delay between the filling of final containers and their labelling and packaging, procedures shall be laid down for the storage of unlabelled containers in order to prevent confusion and to ensure satisfactory storage conditions. Special attention shall be paid to the storage of heat labile or photosensitive products. Storage temperatures shall be specified.

VII.22.A system to ensure the integrity and closure of containers after filling shall be implemented where the final products or intermediates pose specific risks and procedures shall be established to deal with any leaks or spillages. There shall be also procedures in place regarding filling and packaging operations so as to maintain the product within relevant specified limits, e.g. time and/or temperature.

VII.23.The handling of vials (including the capping thereof) containing live biological agents shall be performed so as to prevent the contamination of other products or egress of the live agents into the work environment or the external environment. The viability of such organisms and their biological classification shall be taken into consideration as part of the management of such risks.

VII.24.The suitability of primary packaging materials having regard to the characteristics of the product and the storage conditions (e.g. products that should be stored at ultra-low temperature) shall be ensured. The compatibility of labels with ultra-low storage temperatures, where such temperatures are used, shall be verified.

VIII.   QUALITY CONTROL

VIII.1.As controls for biological products usually involve biological analytical techniques, which typically have a greater variability than physico-chemical determinations, particular attention shall be paid to in-process controls. In-process control testing shall be performed at appropriate stages of production to control those conditions that are important for the quality of the finished product. Particular attention shall be paid to quality controls when continuous culture is used.

VIII.2.Continuous monitoring of data during a production process may be required, for example monitoring of physical parameters during fermentation.

VIII.3.It may be necessary to retain samples of intermediate products in sufficient amount and under appropriate storage conditions to allow repetition or confirmation of a batch control.

VIII.4.Where intermediates can be stored for extended periods of time (days, weeks or longer), consideration shall be given to the inclusion of finished product batches made from materials held for their maximum in-process periods in the on-going stability programme.

VIII.5.The ongoing stability monitoring may require animal testing. In such cases, where no alternative testing methods are available and with a view to reduce the use of animals for testing purposes, the frequency of testing may be adapted under a risk-based approach. Bracketing and matrix approaches may also be applied if scientifically justified in the stability protocol.

VIII.6.For cellular products, sterility tests shall be conducted on antibiotic-free cultures of cells or cell banks to provide evidence for absence of bacterial and fungal contamination and to be able to detect fastidious organisms where appropriate.

VIII.7.For biological medicinal products with a short shelf life (i.e. a period of 14 days or less) requiring batch certification before completion of all end product quality control tests (e.g. sterility tests) a suitable control strategy shall be put in place taking into account the specific characteristics of the product and manufacturing process and taking into account the controls and attributes of starting and raw materials. A detailed description of the release procedure, including the responsibilities of the different personnel involved in assessment of production and analytical data is required. A continuous assessment of the effectiveness of the quality assurance system shall be in place including records kept in a manner that permits trend evaluation.

Where end product tests are not available due to their short shelf life, alternative methods of obtaining equivalent data to permit initial batch certification may be considered (e.g. rapid microbiological methods). The procedure for batch certification and release may be carried out in two or more stages:

— assessment (by a designated person) of batch processing records, the results from environmental monitoring (where available), all deviations from standard procedures and the available analytical results;

— assessment of the final analytical tests and other information available for final certification by the Qualified Person. A procedure shall be in place to describe the measures to be taken where out of specification test results are obtained. Such events shall be fully investigated and the relevant corrective and preventive actions taken to prevent recurrence documented.

IX.   SPECIFIC REQURIMENTS FOR SELECTED PRODUCT TYPES

IX.1.   Allergen products

The following additional requirements are applicable for allergen products:

(a) Source materials shall be described in sufficient detail to ensure consistency in their supply, e.g. common and scientific name, origin, nature, contaminant limits, method of collection. Materials derived from animals shall be from healthy sources. Appropriate biosecurity controls shall be in place for colonies (e.g. mites, animals) used for the extraction of allergens. Allergen products shall be stored under defined conditions to minimise deterioration.

(b) The production process steps including pre-treatment, extraction, filtration, dialysis, concentration or freeze-drying steps shall be described in detail and validated.

(c) The modification processes to manufacture modified allergen extracts (e.g. allergoids (⁴⁹), conjugates) shall be described. Intermediates in the manufacturing process shall also be identified and controlled.

(d) Allergen extract mixtures shall be prepared from individual extracts from single source materials. Each individual extract shall be considered as one active substance.

For recombinant allergens, the additional requirements in Section IV.4 also apply.

IX.2.   Animal immunosera products

The following additional requirements are applicable for animal immunosera products:

(a) Particular care shall be paid to the control of antigens of biological origin to assure their quality, consistency and absence of contamination from adventitious agents. The preparation of materials used to immunise the source animals (e.g. antigens, hapten (⁵⁰) carriers, adjuvants, stabilising agents) as well as the storage conditions for such material immediately prior to immunisation shall be in accordance with documented procedures.

(b) The immunisation, test bleed and harvest bleed schedules shall be in accordance with the terms of the marketing authorisation.

(c) The manufacturing conditions for the preparation of antibody sub-fragments (e.g. Fab or F(ab’)2) and any further modifications shall be in accordance with validated parameters. Where such enzymes are made up of several components, their consistency shall be assured.

IX.3.   Vaccines

The following additional requirements are applicable for vaccines:

(a) Where eggs are used, the health status of all source flocks used in the production of eggs (whether specified pathogen free or healthy flocks) shall be assured.

(b) The integrity of containers used to store intermediate products and the hold times shall be validated.

(c) The sequence of addition of active ingredients, adjuvants and excipients during the formulation of an intermediate or final product shall be in compliance with specifications.

(d) Where the manufacture or testing involves the handling of organisms with a higher biological safety level (e.g. panzootic vaccine strains), appropriate containment arrangements shall be in place in accordance with relevant national requirements. Relevant approvals shall be available for verification.

IX.4.   Recombinant products

The following additional requirements are applicable for recombinant products:

(a) Process conditions during cell growth, protein expression and purification shall be maintained within validated parameters to ensure consistent production with a defined range of impurities. Depending on the type of cell used in production, additional measures may be required to ensure viral safety. When the manufacturing process involves multiple harvests, the period of continuous cultivation shall be within specified limits.

(b) The purification processes to remove unwanted host cell proteins, nucleic acids, carbohydrates, viruses and other impurities shall be within defined validated limits.

IX.5.   Monoclonal antibody products

The following additional requirements are applicable for monoclonal antibody products:

(a) Monoclonal antibodies may be manufactured from hybridomas or by recombinant DNA technology. Control measures appropriate to the different source cells (including feeder cells if used) and materials used to establish the hybridoma/cell line shall be in place to assure the safety and quality of the product. It shall be verified that these are within approved limits. Viral safety is particularly important. Data originating from products generated by the same manufacturing technology platform may be acceptable to demonstrate suitability.

(b) Production and product parameters at the end of a production cycle (e.g. temperature, Ph, density, oxygen, cell viability, etc.) and for early termination of production cycles shall be defined and monitored.

(c) The manufacturing conditions for the preparation of antibody sub-fragments (e.g. Fab, F(ab’)2, scFv) and any further modifications (e.g. radio labelling, conjugation, chemical linking) shall be in accordance with validated parameters.

IX.6.   Veterinary medicinal products derived from genetically modified/genome edited animals

The following additional requirements are applicable for veterinary medicinal products derived from genetically modified/genome edited animals:

(a) Animals used for production shall be clearly and uniquely identified and backup arrangements shall be put in place in the event of loss of the primary marker.

(b) The genealogy of the founder animals through to production animals shall be documented. Since the genetically modified/genome edited line will be derived from a single genetic founder animal, materials from different genetically modified/genome edited lines shall not be mixed.

(c) The conditions under which the product is harvested shall be in accordance with the terms of the marketing authorisation. The harvest schedule and conditions under which animals may be removed from production shall be performed according to approved procedures and acceptance limits.

(d) Particular attention shall be made to demonstration of batch-to-batch consistency.

IX.7.   Veterinary medicinal products derived from genetically modified/genome edited plants

The following additional requirements are applicable for veterinary medicinal products derived from genetically modified/genome edited plants.

(a) Additional specific measures may be required to prevent contamination of the master and working genetically modified/genome edited banks by extraneous plant materials and relevant adventitious agents. The stability of the gene within the defined generation numbers shall be monitored.

(b) Plants shall be clearly and uniquely identified, the presence of key plant features, including health status, across the crop shall be verified at defined intervals through the cultivation period to assure consistency of yield between crops.

(c) Security arrangements for the protection of crops shall be defined, wherever possible, to minimise the exposure to contamination by microbiological agents and cross-contamination with non-related plants. Measures shall be put in place to prevent materials such as pesticides and fertilisers from contaminating the product. A monitoring programme shall be established and all results documented. Any incident shall be investigated and its impact on the continuation of the crop in the production programme shall be determined.

(d) Conditions under which plants may be removed from production shall be defined. Acceptance limits shall be set for materials (e.g. host proteins) that may interfere with the purification process. It shall be verified that the results are within the approved limits.

(e) Environmental conditions (temperature, rain) that may affect the quality attributes and yield of the recombinant protein from the time of planting, through cultivation to harvest and interim storage of harvested materials shall be documented. The principles in documents such as ‘Guideline on Good Agricultural and Collection Practice for Starting Materials of Herbal Origin’ (⁵¹) shall be taken into account when drawing up such conditions.

(f) Particular attention shall be made to the demonstration of batch-to-batch consistency.

ANNEX III

This Annex provides for additional requirements and specific adaptations to the requirements set out in this Regulation which shall apply to certain types of veterinary medicinal products.

Unless stated otherwise, the requirements contained in this Annex shall apply in addition to the requirements provided for in the Regulation. In case of conflict, the specific requirements set out in this Annex shall prevail.

I. HERBAL VETERINARY MEDICINAL PRODUCTS

I.1.Taking into account the variability of herbal materials, the control of herbal materials (herbal substances and herbal preparations) used in the manufacture of veterinary medicinal products is particularly important.

I.2.Herbal materials used in the manufacture of veterinary medicinal products shall be of suitable quality. The selection of seeds, cultivation and harvesting conditions are important aspects of the quality of the herbal substance that can influence the consistency of the finished product.

I.3.Table 1 illustrates the application of good practices in connection with the manufacture of veterinary medicinal products.

Activity	Good agricultural and collection practice	GMP for active substances used in the manufacture of veterinary medicinal products or GMP for veterinary medicinal products, as applicable	GMP for veterinary medicinal products
Cultivation, collection and harvesting of plants, algae, fungi and lichens, and collection of exudates	Applicable
Cutting, and drying of plants, algae, fungi, lichens and exudates (1)	Applicable	Applicable
Expression from plants and distillation (2)		Applicable
Comminution, processing of exudates, extraction from plants, fractionation, purification, concentration or fermentation of herbal substances		Applicable
Further processing into a dosage form, including packaging as finished veterinary medicinal product			Applicable
(1) Manufacturers shall carry out these steps in accordance with the marketing authorisation. For initial steps that take place in the field, as justified in the marketing authorisation, the standards of good agricultural and collection practice for starting materials of herbal origin (GACP) shall apply. Good manufacturing practice shall apply to further cutting and drying steps. (2) Regarding the expression from plants and distillation, if it is necessary for these activities to be an integral part of harvesting to maintain the quality of the product within the approved specifications, it is acceptable that they are performed in the field, provided that the cultivation is in compliance with GACP. This approach may only be accepted in exceptional cases and provided that it is agreed in the relevant marketing authorisation. For activities carried out in the field, appropriate documentation, control and validation according to good manufacturing principles shall be assured.

I.4.The herbal materials used in the manufacture of veterinary medicinal products shall comply with the following:

(a) Specifications shall be set in compliance with the marketing authorisation and shall include: — the binomial scientific name of plant (genus, species, subspecies/variety and author (e.g. Linnaeus); other relevant information such as the cultivar name and the chemotype shall also be provided, as appropriate; — details of the source of the plant (country or region of origin, and where applicable, cultivation, time of harvesting, collection procedures, possible pesticides used, possible radioactive contamination, etc.); — which part(s) of the plant is/are used; — when a dried plant is used, the drying system shall be specified; — a description of the herbal substance and its macro and microscopic examination; — suitable identification tests including, where appropriate, identification tests for constituents with known therapeutic activity or markers. Specific distinctive tests are required where an herbal substance is liable to be adulterated/ substituted. A reference authentic specimen shall be available for identification purposes; — the water content for herbal substances, which is to be determined in accordance with the European Pharmacopoeia; — assay of constituents of known therapeutic activity or, where appropriate, of markers; — methods to determine a possible pesticide contamination and the accepted limits, in accordance with European Pharmacopoeia methods or, in the absence thereof, with an appropriate validated method, unless otherwise justified; — tests to determine fungal and/or microbial contamination, including aflatoxins, other mycotoxins, pest-infestations and limits accepted, as appropriate; — tests for toxic metals and for likely contaminants and adulterants, as appropriate; — tests for foreign materials, as appropriate; — any other additional test according to the European Pharmacopoeia general monograph on herbal substances or to the specific monograph of the herbal substance, as appropriate. Any treatment used to reduce fungal/microbial contamination or other infestation shall be specified. Specifications shall include details of the process, tests and limits for residues, as appropriate.

(b) Suppliers of herbal materials shall comply with good agricultural and collection practice. The suppliers shall be audited by the manufacturer of the herbal veterinary medicinal product in accordance with quality risk management principles. Such audits may be outsourced.

I.5.The following precautions shall be taken regarding storage areas for herbal materials used in the production of veterinary medicinal products:

(a) effective measures shall be implemented to prevent the spread of insects, other animals or micro-organisms brought in with the herbal substance, to prevent fermentation or mould growth and to prevent cross-contamination. Separate enclosed areas shall be used to quarantine incoming herbal substances and for the approved herbal substances;

(b) storage areas shall be well aerated, and the containers shall be located in such a way so as to allow free circulation of air;

(c) where necessary, specific conditions of humidity, temperature or light protection shall be defined and monitored.

I.6.The identity and quality of herbal materials and of herbal medicinal products shall be determined in accordance with the relevant current European guidance on quality and specifications for herbal medicinal products and traditional herbal medicinal products or, where relevant, with the requirements of specific relevant monographs of the European Pharmacopoeia.

I.7.The processing instructions shall describe the different operations to be carried out upon the herbal substance such as cleaning, drying, crushing and sifting, including drying time and temperatures, and methods used to control the cut size or the particle size. Written instructions shall be developed and records shall be kept to ensure that each container of herbal substance is carefully examined to detect any adulteration/substitution or presence of foreign matter, such as metal or glass pieces, animal parts or excrement, stones, sand, or rot and signs of decay.

The processing instructions shall also describe security sieving or other methods of removing foreign materials and appropriate procedures for the cleaning/selection of the plant material before the storage of the approved herbal substance or before the start of manufacturing.

For the production of an herbal preparation, instructions shall include details of solvent, time and temperature of extraction, details of any concentration stages and methods used.

I.8.If dust is generated during processing (including sampling), the use of dust extraction, dedicated premises or other means shall be considered to prevent cross-contamination and facilitate cleaning.

I.9.The equipment and filtering materials used in the manufacturing process shall be compatible with the extraction solvent, in order to prevent any release or undesirable absorption of the substance that could affect the product.

I.10.Due to the fact that medicinal plant/herbal substances are heterogeneous in nature, the following measures shall be implemented regarding sampling:

(a) each batch shall be identified by its own documentation;

(b) a reference sample of the plant material is necessary, especially in those cases where the herbal substance is not described in the European Pharmacopoeia or in another Pharmacopoeia of a Member State. Samples of unmilled plant material are required if powders are used.

I.11.Quality control personnel shall have particular expertise and experience in herbal substances, herbal preparations and/or herbal medicinal products in order to be able to carry out identification tests and recognise adulteration, the presence of fungal growth, infestations, non-uniformity within a delivery of crude material, etc.

II.   VETERINARY MEDICINAL PRODUCTS INTENDED FOR INCORPORATION INTO MEDICATED FEEDINGSTUFFS

II.1.Because the manufacture of medicated premixes requires the use of large quantities of vegetable matter which is likely to attract insects and rodents, it is particularly important to ensure that premises are designed, equipped and operated in a way that minimises the risk of intrusion thereof in the site. Enhanced pest control systems shall be put in place to monitor and minimise pest ingress and to take action where appropriate.

II.2.Because of the large volume of dust generated during the production of bulk material for premixes, specific attention shall be paid to the need to avoid cross contamination and facilitate cleaning, for example through the installation of sealed transport systems and dust extraction, whenever possible. The installation of such systems does not, however, eliminate the need for regular cleaning of production areas.

II.3.Parts of the process likely to have a significant adverse influence on the stability of the active ingredient(s) (e.g. use of steam in pellet manufacture) shall be carried out in an uniform manner from batch to batch.

II.4.Whenever possible, the manufacture of premixes shall be made in dedicated areas which, if possible, do not form part of the main manufacturing plant. Alternatively, such dedicated areas shall be surrounded by a buffer zone in order to minimise the risk of contamination of other manufacturing areas.

III.   ECTOPARASITIC VETERINARY MEDICINAL PRODUCTS

Ectoparasitic veterinary medicinal products for external application to animals may be produced and filled on a campaign basis in pesticide specific areas. However, other categories of veterinary medicinal products shall not be produced in such areas.

IV.   LIQUIDS, CREAMS AND OINTMENTS

As liquids, creams and ointments may be particularly susceptible to microbial and other contamination during manufacture, the following measures shall be considered:

(a) the use of closed systems for processing and transfer is recommended. In cases where the products or open clean containers are exposed to the environment, there shall be effective ventilation with filtered air;

(b) tanks, containers, pipework and pumps shall be designed and installed to facilitate cleaning. In particular, equipment shall include a minimum of dead-legs or sites where residues can accumulate and promote microbial proliferation;

(c) the use of glass apparatus shall be avoided wherever possible. High quality stainless steel is often the material of choice for parts coming into contact with the product;

(d) the chemical and microbiological quality of water used in production shall be specified and monitored. Care shall be taken in the maintenance of water systems in order to avoid the risk of microbial proliferation. After any chemical sanitisation of the water systems, a validated flushing procedure shall be applied to ensure that the sanitising agent has been effectively removed;

(e) materials likely to shed fibres or other contaminants, like cardboard or wooden pallets, shall not enter the areas where products or clean containers are exposed;

(f) the homogeneity of mixtures or suspensions, shall be kept during filling. Special care shall be taken at the beginning of a filling process, after stoppages and at the end of the process to ensure that homogeneity is maintained;

(g) when the finished product is not immediately packaged, the maximum period of storage and the storage conditions shall be specified and adhered to.

V. MEDICINAL GASES

V.1.   Scope

This Section provides for additional requirements that are applicable to the manufacture of veterinary medicinal products that contain medicinal gases. For the purposes of this Section, the term ‘gas’ covers any substance that is completely gaseous at 1,013 bar and + 20 °C or has a vapour pressure exceeding 3 bar at + 50 °C.

In the exceptional case of continuous manufacturing, where no intermediate storage of the gas between the manufacture of the active substance and the manufacture of the medicinal product is possible, the whole process (from starting materials of active substance to medicinal finished product) falls under the scope of this Regulation.

V.2.   Personnel

Personnel shall be specifically trained on the specific hazards from these products; training programs shall include tanker lorries drivers and subcontracted personnel that can influence the quality of medicinal gases (such as personnel in charge of maintenance of cylinders (⁵²) or valves).

V.3.   Premises

V.3.1.Cylinders and mobile cryogenic vessels (⁵³) shall be checked, prepared, filled and stored in separate areas from non-medicinal gases, and there shall be no exchange of cylinders/mobile cryogenic vessels between these areas. However, it is acceptable to check, prepare, fill and store other gases in the same areas, provided that such operations are performed according to good manufacturing practice.

V.3.2.Premises shall be designed to provide separate marked areas for different gases and clear identification and segregation of cylinders/mobile cryogenic vessels at various stages of processing (e.g. ‘waiting checking’‘awaiting filling’, ‘quarantine’, ‘certified’, ‘rejected’, ‘prepared deliveries’). The method used to achieve these various levels of segregation will depend on the nature, extent and complexity of the overall operation. Marked-out floor areas, partitions, barriers, signs, labels or other appropriate means may be used.

V.3.3.Cylinders/home cryogenic vessels (⁵⁴) (whether empty after sorting or maintenance or filled) shall be stored under cover and protected from adverse weather conditions. Filled cylinders/mobile cryogenic vessels shall be stored in a manner that ensures that they will be delivered in a clean state, compatible with the environment in which they will be used.

V.3.4.Specific storage conditions required by the marketing authorisation (e.g. for gas mixtures where phase separation occurs on freezing) shall be respected.

V.4.   Equipment

V.4.1.Equipment shall be designed to ensure that the correct gas is filled into the correct container. (⁵⁵) There shall normally be no cross connections between pipelines carrying different gases. If cross connections are needed (e.g. filling equipment of mixtures), as part of the qualification process, it shall be ensured that there is no risk of cross contamination between the different gases. In addition, manifolds (⁵⁶) shall be equipped with specific connections. The use of connections meeting different standards at the same filling site shall be carefully controlled, as well as the use of adaptors that may be needed to bypass the specific fill connection systems.

V.4.2.Tanks (⁵⁷) and tankers (⁵⁸) shall be dedicated to a single and defined quality of gas. However, medicinal gases may be stored or transported in the same tanks/containers for intermediate storage/tankers as the same non-medicinal gas, provided that the quality of the latter is at least equal to the quality of the medicinal gas, that standards of good manufacturing practice are maintained and that the approach is justified in accordance with quality risk management principles.

V.4.3.A common system supplying gas to medicinal and non-medicinal gas manifolds is only acceptable if there is a validated method to prevent backflow from the non-medicinal gas line to the medicinal gas line.

V.4.4.Filling manifolds shall be dedicated to a single medicinal gas or to a given mixture of medicinal gases. In exceptional cases, filling gases used for non-medicinal purposes on manifolds used for medicinal gases may be acceptable if duly justified and performed under control. In these cases, the quality of the non-medicinal gas shall be at least equal to the required quality of the medicinal gas and standards of good manufacturing practice shall be maintained. Further, in such cases, the filling shall be carried out on campaign basis.

V.4.5.Repair and maintenance operations (including cleaning and purging (⁵⁹)) of equipment, shall not adversely affect the quality of the medicinal gases. In particular, procedures shall describe the measures to be taken after repair and maintenance operations involving breaches of the system’s integrity. Specifically, it shall be demonstrated that the equipment is free from any contamination that may adversely affect the quality of the finished product before releasing it for use. Records shall be maintained.

V.4.6.A procedure shall describe the measures to be taken when a tanker is back into medicinal gas service (after transporting non-medicinal gas under the conditions referred in Section V.4.2, or after a maintenance operation), which shall include appropriate analytical testing.

V.5.   Documentation

V.5.1.Data included in the records for each batch of medicinal gases shall ensure that each filled container is traceable to significant aspects of the relevant filling operations. As appropriate, the following shall be entered:

— name of the product;

— batch number;

— date and time of the filling operation;

— identification of the person(s) carrying out each significant step (e.g. line clearance, receipt, preparation before filling, filling, etc.);

— batch(es) reference(s) for the gas(es) used for the filling operation, including status;

— equipment used (e.g. filling manifold);

— quantity of cylinders/mobile cryogenic vessels before filling, including individual identification references and water capacity(ies);

— pre-filling operations performed;

— key parameters that are needed to ensure correct filling at standard conditions;

— results of the appropriate checks to ensure that the cylinders/mobile cryogenic vessels have been filled;

— a sample of the batch label;

— specification of the finished product and results of the quality control tests (including reference to the calibration status of the test equipment);

— quantity of rejected cylinders/mobile cryogenic vessels, with individual identification references and reasons for rejections;

— details of any problems or unusual events, and signed authorisation for any deviation from filling instructions;

— certification statement by the qualified person, date and signature.

V.5.2.Records shall be maintained for each batch of gas intended to be delivered into tanks in healthcare facilities. These records shall, as appropriate, include the following:

— name of the product;

— batch number;

— identification reference for the tank;

— date and time of the filling operation;

— identification of the person(s) carrying out the filling of the tank (tanker);

— reference to the supplying tanker (tank), reference to the source gas as applicable;

— relevant details concerning the filling operation;

— specification of the finished product and results of the quality control tests (including reference to the calibration status of the test equipment);

— details of any problems or unusual events, and signed authorisation for any deviation from filling instructions;