VAL-240 Method Validation Master Plan

Table 1: Risk Matrix (Methods)

At the completion of the assessment process the Method Validation assessment is updated (refer Section C) and the Method Validation Master Schedule is also updated (refer Section D). 

4.3.5. New or Changed Systems

[Enter Business name] shall endeavour to conduct method validation activities in a “Prospective” manner where ever possible but concurrent with product development is acceptable. Legacy method validation will be performed for assays in use as defined under Section A, point 2 of this MVMP. 

The change control program (refer to QMS-125 Change Management System) ensures that validated/qualified test methods and laboratory equipment remain in a validated/qualified state.  It is essential to ensure that changes to the equipment, test method and support systems do not affect the validation/qualification status of the test method. If a change is made to qualified equipment or a validated test method, re-qualification or re-validation studies may be required.  

In cases where it is necessary to confirm that a minor change has no effect upon qualified equipment or a validated test method, a confirmation assay may be performed to provide data that may be used to support this hypothesis and close-out the change authorization. The need for re-qualification or re-validation is subject to the requirements of the change control program. 

5. Validation Activities

Validation studies will generally follow the plans outlined in the following sub-sections.

5.1. Equipment Qualification (EQ)

Laboratory and in-process control test equipment qualification is conducted to provide documented evidence that instruments and equipment have been installed and function according to specifications.  For detail on the approach to equipment qualification refer to the Site Validation Master Plan PV-009.

5.2. Method Validation (MV)

Method Validation provides assurance of the accuracy and reliability of the test method for its intended use.  Equipment used for method validation must be operated within specifications, and be adequately calibrated, maintained, and if applicable qualified

Method Validation will be required for the following:

 – Introduction of a new test method or transfer of a test method from one facility to another.

 – Major modification to existing test method as determined through the change management system.

 – Re-registration of a legacy method

Test Method Validation documents are developed executed and approved in accordance with step 7.0 of this plan. Generally, a test method is considered to be validated when pre-defined acceptance criteria have been met. Detail on specific considerations for method validation are included in section 6.0.

5.3. Decommissioning

Decommissioning will be performed to retire equipment and test methods in a controlled manner. Decommissioning will be triggered through the site change management system. 

5.4. Validation Approaches

Test Method Validation approaches can be prospective or concurrent. Test Method Validation on a new or transferred assay must be successfully completed prior to the QA release and distribution of any product manufactured using that test method. Completion is defined as approval of the summary report documenting compliance with all established acceptance criteria.

Concurrent validation of a test method can only be performed if it is validation of an existing legacy test method or done in parallel with development of a new product. Concurrent validation studies will be approved by Quality.

Method development / optimisation (also referred to as commissioning) may occur before method validation. 

5.5. Pre-requisites for Method Validation

Prior to execution of a Test Method Validation study, the following must be addressed, whenever applicable: 

 – The test method involved must be defined, including operational ranges, critical test parameters, and critical steps.  i.e test method development and optimization must be completed and final reports approved (or in the case of a legacy assay the standard test procedure must be defined).

 – The Method Validation Plan/Protocol must be developed and approved; specifying predetermined and supported acceptance criteria for each experiment prior to the initiation of the method validation.

 – Equipment used in the validation studies must be qualified in compliance with the site VMP.

 – Critical utilities used in the validation studies must be qualified in compliance with the site VMP.  

 – Computer systems used in the validation studies must be qualified in compliance with the site VMP (though it is noted that some aspects of computer system validation for test equipment may be inextricably linked to validation of the test method and will therefore be validated at the same time the method is validated)

 – Critical instrumentation must be calibrated.

 – Personnel involved with the execution of the validation study must be properly trained in the relevant procedures and validation protocol sections.

5.6. General Acceptance Criteria for Method Validation

The following general acceptance criteria should be evaluated for applicability for each method validation study:

 – Acceptance criteria are chosen so as to be both necessary and sufficient to assess the validation of the Method. As much as possible, they should be measurable and not be procedural.  If applicable, source documentation for acceptance criteria must be referenced.  

 – The acceptance criteria should encompass the requirements and specifications used to evaluate the validity of the method during day to day use.

 – Intra-batch (i.e variability between different samples and / or reagents) and inter-assay (i.e. variability between same sample in separate assays) variability should be assessed whenever applicable.

 – Deviations related to the execution of validation studies must be reported in a timely manner and, if necessary, investigated to determine the root cause.  These deviations must be documented and resolved according to the deviation reporting procedure within the protocol.

As previously indicated, each validation study should be summarized in an approved report.

5.7. Validation & Qualification Projects

Current validation projects are noted in the Schedule of Test Method Validation Activities, refer section D.  

6. Specific Considerations for Method Validations  

The following are specific considerations that are to be taken into account when validating methods or receiving test methods as part of a technology transfer. Not all considerations apply to all methods; the specific test method validation protocol and/or summary report should justify the selection of validation criteria. The methods intended use and type would drive this selection process.

Reference should be made to the following guidelines:

 – Guideline on Validation of Analytical Procedures: Definition and Terminology [VICH Topic GL1]

 – Guideline on Validation of Analytical Procedures: Methodology [VICH Topic GL2]

 – Note for Guidance on Validation of Analytical Procedures: Text & Methodology [ICH Topic Q2 (R1)] when determining the test validation approach.

A matrix approach is acceptable for method validation where justified. E.g. 5 products A, B, C, D and E may require sterility testing, however due to the amount of preservative product C is considered worst case. Product C will be used in the validation.

6.1. Analytical Methods

An analytical assay is defined as an assay that gives a quantitative result and is measuring a known entity. This may be a chemical assay or an immunological assay. Tests to determine the following should be performed at least three times.

Specificity: Ability to assess unequivocally the analyte in the presence of component’s which may be expected to be present.  

Accuracy: Closeness of agreement between the true value and the value found.

Precision: Closeness of agreement (degree of scatter) between a series of measurements obtained from multiple sampling of the same homogeneous sample.

Detection: Limit Lowest amount of analyte in a sample that can be detected but not necessarily quantitated as an exact value.

Quantitation: Limit Lowest amount of analyte in a sample that can be detected with suitable accuracy and precision.

Linearity:  Ability (within a given range) to obtain test results that are directly proportional to the concentration of analyte in sample.

Range: The interval between lower and upper concentrations (amounts) of analyte sample that can be measured with a suitable accuracy, precision and linearity. 

Ruggedness: Degree of reproducibility of test results under varying conditions, such as different operator, instruments, reagents, etc.

Robustness: Capacity to remain unaffected by deliberate changes and / or variations in method parameters. (reliability and stability)

 6.2. Compendia Based Methods

For [Enter Business name] compendial methods currently include recognised compendia such as the EP, BP and USP. In addition, for the purpose of defining validation requirements, test methods published by the TGA / USDA (i.e. 9CFR.) or other government bodies and the AOAC are considered the equivalent to compendia methods and, therefore, fall under the definition of a compendia-based method.

Compendia methods are considered validated but must be verified under actual conditions of use.  The suitability of a compendia analytical method is verified by: 

 – Meeting the requirements for validation if stated in the compendia monograph.

 – Meeting the requirements of controls in the compendia monograph.

 – Demonstrating appropriate specificity (e.g., there is no interference in the method from any impurities, degradants, or excipients for the product).

 – Demonstrating the ability to run the compendia method under actual conditions of use.

Changes to existing compendia methods may occur. An evaluation of the change(s) should occur, and recommendations should be made in regard to local activities and the need for verification testing.

6.3. Microbiological Methods

6.3.1. General 

Sterility, non-viability and preservative effectiveness are the most common microbiological test methods used to determine vaccine quality. Methods should be validated against compendial requirements which often state the validation requirements.

It is noted that for microbiological assays the validation is often part of the assay development e.g. non-viability assay and therefore a separate validation post method development is not always required.

For sterility testing, this normally means demonstrating that the test media chosen can support the growth of various challenge microorganisms in the presence of the product and that the product is not growth inhibitory.

For non-viability testing this normally means demonstrating that the test media chosen is the most sensitive for the specific micro-organism that the inactivated product is derived from. This is generally a comparison between the manufacturing media or another recognized media that provides good growth conditions for the microorganism and that there is no interference to growth from the residual inactivant. During the test validation, a comparison between solid, liquid and subculturing should be considered.

Preservative effectiveness testing must be performed per compendial requirements. Microorganisms used for this test are determined by compendial requirements.  Other strains may be included, based upon formulation and intended use.  The inclusion of environmental isolates needs to be considered.

Alternative microbiological procedures, including automated methods, may be used, provided that they have been validated and have been proven to be equivalent or superior to the compendial or existing method.

6.3.2. Test Design Considerations

Compendial requirements should be taken into account when designing a microbiological assay. The dilution (test sensitivity) at which a sample is plated must be sensitive enough to allow verification that the specification for that test sample has been met.  

Where cultures are used, they should be traceable, and preferably be prepared fresh (≤ 24 hours). Microbial cultures (i.e., suspensions/lyophilized) purchased from an approved vendor may be used directly from the container/vial. When determining the growth promotion organisms to be used testing must be performed in accordance with compendial requirements.  When not defined by compendia, select cultures and incubation parameters according to specific test expectations.  

The incubation periods in validation protocols should be specified. 

6.3.3 Validation Considerations

Validation of microbiological assay is generally performed differently to that of an analytical assay. A microbiological assay is qualified using test organisms to demonstrate growth promotion, to demonstrate a lack of test sample interference or if a preservative is present to demonstrate interference.

Analytical microbiological assays where used (e.g. Thiomersal concentration determination using zones of inhibition) are validated following the guidelines of any analytical test. Dilutions and transfers must be performed in a quantitative manner and the layout of discs (or equivalent) for diffusion assays must have discreet zones of inhibition.

A compendial assay, if explicit in the method and if there are no stated validation requirements does not need validating so long as the method is followed exactly. Any differences require validation. Verification under use is required. 

A compendial assay, if there are stated validation requirements must be validated as per the stated compendial requirements.

The number of replicates for microbiological assay validation is to be enough to give confidence that the assay is reproducible and is to be justified in protocol, the number of replicates can be reduced if ongoing qualification (e.g. use of challenge organisms in the presence of product) forms part of the standard method procedure.

All media that is sterilised must be sterilised using validated load patterns and autoclaves. New media must be assessed to determine if it fits within the existing validation for size, load and container closure. If the new media is supported, then only growth promotion validation is required.

To validate shelf life of media growth promotion studies of test media should be performed at time of preparation and/or receipt and at expiry date. Melting cycles, and molten media hold times must be validated (e.g. temperature, time limits) or growth promotion must be performed post the melting cycle during validation and for routine quality control.

6.4. Animal Test Methods

Animal test methods fall into five main categories; 

 – safety (also known as toxicity) testing, 

 – Challenge testing,

 – Antigen potency assessment 

 – Serum generation, and

 – antibody assessment. 

A compendial assay, if explicit in the method and if there are no stated validation requirements does not need validating so long as the method is followed exactly. Any differences require validation. Verification under use is required. Verification requirements are to be defined under the method protocol.

A compendial assay, if there are stated validation requirements must be validated as per the stated compendial requirements.

When validating an animal based method, the extent of the validation required must be balanced with the animal welfare cost. Reference to scientific literature can be leveraged to avoid unnecessary duplication of in-vivo testing.

Safety or toxicity tests require injecting a certain dose of product into an animal and determining if the product is safe or toxic / non-toxic. Validation considerations would include ability to discern between toxic / non-toxic (or safe / unsafe) material, dilution variation (if applicable) and dose application variation, (Intravenous, Sub-cut, Intraperitoneal). Due to the high animal welfare cost of validating these assays, if a risk based assessment has been written (and approved by QA) documenting the logic of the test, the likelihood of a false pass and field impact and an argument not to validate then test will not be validated.

Challenge testing, by its very nature, involves a set of vaccinates and a set of un-vaccinated control animals. Simplistically, vaccinates must survive the challenge and control animals must die. As the method has built in controls that qualify the assay each time it is performed and the method has high animal welfare cost, challenge tests are generally not validated.

Antigen potency testing involves an unknown amount of toxin or toxoid that is combined with known amounts and values of antitoxin or antitoxin / toxin combination at various levels. An indicator is added which may be in-vitro (e.g. blood) or in-vivo (e.g. mice) on which excess toxin can exhibit its effect. These assays are considered immunological analytical assays and should be validated following the same principles used to validate any analytical method. NB: Based on a documented justification comparing benefit to animal welfare cost for in-vivo assays not all aspects may need validating.

Serum generation is part of a potency assay. The timing of maximum antibody generation in a test animal is normally not critical as the pass value for the product is assigned from the vaccination schedule and is linked to product efficacy trials or compendial instructions. The QC method must reflect the same timing period. The vaccination schedule can impact on whether there is a linear dose response in the animal. The vaccination schedule should be validated to demonstrate that a linear dose response occurs and that a sub potent vaccine can be differentiated from an efficacious vaccine.

In-vivo antibody assessment methods are immunological analytical assays and should be validated following the same principles used to validate any analytical method. NB: Based on a documented justification comparing benefit to animal welfare cost for in-vivo assays not all aspects may need validating.

6.5. Legacy Test Methods

Legacy test methods are defined as those methods in current use prior to the approval of the original of this MVMP and where a validation package is known not to exist. Validation of a legacy test methods follows the same principles as validation of new methods however test performance data may be extracted from existing historical records rather than repeating assays.

The validation scope for a legacy assay may be reduced based on a documented justification and review of the historical performance of the assay and assessment of known assay.

Legacy compendia methods (as defined in the MVMP) are deemed verified based on an history of successful use.

6.6. Technology Transfers

Qualification of the receiving laboratory must be completed before the test procedure is used to release commercial product. Management is to identify transfer team members from the participating laboratories (e.g., receiving laboratory and transferring laboratory). The transfer team members are responsible for managing the transfer and qualification process and to ensure that all activities meet the requirements of this MVMP.

The results of the validation and transfer experiments shall meet predetermined acceptance criteria to be considered acceptable.  The following principles for test transfer to be followed: 

– An analytical package is prepared by the transferring laboratory which includes, but is not limited to:

 – Test procedure(s) for which qualification is required.

 – Test procedure development and validation reports.

 – Proposed specifications/controls applicable to the test procedure(s). NB: For legacy assays these may not be known and will need developing as part of the transfer.

– Information supplied is to be reviewed by the receiving site. During the review stage, a checklist is created that itemizes all requirements of the test procedure (including, but not limited to, people (skills/experience needed), equipment, facilities, and local site procedures) that must be evaluated during the receiving laboratory readiness assessment.

– From this review a gap analysis is performed and an action plan for achieving receiving laboratory readiness must be completed prior to starting the qualification phase. A strategy document should then describe in detail, the following:

 – List of analytical test procedures and equipment that require qualification.

 – Training plan outlining method familiarization (if required) and laboratory trial requirements.

 – Plan outlining the qualification option selected, with justification, for each test procedure to be transferred.

– Sharing of lab note books and records should occur during this phase.

– Transfer Process – Qualification. The qualification phase consists of documenting the training process and demonstrating performance qualification against a pre-approved qualification protocol, and contains pre-defined acceptance criteria for data comparisons and suitability of method performance. Training process consisting of the following:

 – Method familiarisation typically involves the trainee from the receiving laboratory working with a qualified analyst from the transferring laboratory to perform a review of the test procedure, observe the equipment, and review any associated safety procedures.  If necessary, the qualified analyst will perform all laboratory techniques required to perform the test procedure.

 – Method familiarisation can take place at either the transferring or receiving laboratory.  If the basis of the method is familiar and fits within existing test methodology familiarization can be done verbally. The familiarisation methods will be determined by the Transfer Team (e.g., teleconference between locations may occur) depending on the project.

 – Lab demonstration trials are performed to demonstrate that the necessary test procedure techniques, equipment, facility operation, and safety procedures relating to the analytical technology can be applied in the receiving environment.

Qualification Phase may utilise one or more or a combination of five options to qualify analysts at the receiving laboratory.  A summary of the five Qualification Phase options is provided below: 

 – Comparative Testing qualification is the demonstration that when the trainee prepares specified samples the equipment is satisfactorily operating for its intended use in the test procedure (such as when performing linearity, sensitivity experiments, etc.).  The design of experiments should allow the comparison of results generated on a reference test article, where possible, by the trainee at the receiving laboratory to results generated on the same representative test article by a qualified analyst at the transferring laboratory. These test articles can be new samples, previously analysed samples or blind samples provided to both laboratories.

 – Test Procedure Validation is used when results from performing key elements of the test method validation by the trainee in the receiving laboratory are compared and must be consistent with previously demonstrated method capabilities and validation results.

 – Test Procedure Verification of Compendial Methods is used to qualify a method when results are verified by establishing that the method meets the requirements of the monograph at the receiving laboratory.

 – Experience with a Highly Similar Test Procedure can be used as justification of qualification when the receiving laboratory is using methodology that is scientifically similar to the test procedure which is already being performed by qualified analysts in the receiving laboratory.

 – Experience with a Highly Similar Procedure may be applied to test procedures for potency and purity of the active ingredient when the following conditions are satisfied: (1) The test procedures have equivalent instrumental requirements and (2) The test article sample preparations are comparable.

7. Validation documentation 

The [Enter Business name] Validation program uses two types of controlled documentation, System management or guidance documentation and evidence of compliance documentation. The level of documentation required will be defined based on the type of study, potential risk and complexity of the study.

7.1. System Management Documentation (SM)

System Management documents are defined as those documents that describe the management of a Validation system or those documents created to provide guidance on critical tasks (e.g. method validation master plan).

7.2. Evidence of Validation documents 

Evidence of Validation documents are those documents that provide a level of assure that a method has been validated and is fit for its intended use. 

 – Validation protocols are developed specifying instructions for execution and pre-determined acceptance criteria. Studies are executed in conformity with the validation protocol. The objective of each protocol is to provide clear and complete instructions for the execution of a validation study. 

 – Validation data is collected, recorded, analyzed, and summarized with a conclusion in a Validation Summary Report.  Any unexpected events encountered during the execution are documented and thoroughly investigated. Investigation reports are reviewed and approved prior to the approval of the Summary Report. 

Assays may be validated in Process Development or R&D departments. Protocols follow scientific format and the outcome of each protocol is a scientific report. As test methods may be validated off site by other [Enter Business name] bodies (e.g. International R&D) documentation may differ in appearance from that used on site.

PART C – Listing and Qualification Status of Methods in Use

8. Qualification Status of Methods

Part C of this document, entitled Listing and Qualification Status of methods in use provides a full list of test methods in use and their current, historical validation state.  Each method has been risk assessed in accordance with this plan.  Part C is a stand alone, approved document.

PART D – Schedule of Validation Activities 

9. Test Validation Activities

Part D of this document is a schedule of planned test validation activities. 

The schedules provide the current status of test method validation. The schedule will be updated on an annual basis by the QC Manager(s). Part D is a stand-alone, approved document.

10. Revision history