Assay Qualification/Validation – a Reviewer’s Expectations

1 Assay Qualification/Validation . a Reviewer's Expectations Sarah Kennett Division of Monoclonal Antibodies Office of Biotechnology Products OPS/CDER/FDA. 1. October 20, 2010. Disclaimer The views and opinions expressed here are my own and should not be used in place of regulations, published FDA guidances or discussions with the Agency. These views reflect CDER/OPS/OBP. discussions. This talk is focused on analytical assays used to assess drug substance, drug product, and in-process materials. 2. Overview Why this subject? What does FDA say officially? What do we OBP product reviewers mean by qualification and validation ? Some Expectations 3. Person X: Someone told me assays need to be validated to start Phase 2 studies. Me: No, unless there is a scientific reason, submission of Assay validation isn't required until the BLA is submitted. However, you need to be using qualified assays from the beginning.

2 If the assays you're using during later phases of clinical development can't be validated, you might be in trouble regarding setting of acceptance criteria and dosing/efficacy assessment. X: So, we need to show you validated assays by Phase 3. Me: Uhh, no, not exactly. Formal validation studies CAN be performed as early as you want, but with a few exceptions, the REQUIREMENT regarding timing of the original validation {exceptions include assays critical for safety!} is that it be completed prior to submission of the BLA to be included in the submission. For phase 3, you probably need to be using assays that can be validated, though, so they need to at least be well qualified. X: 4. Me: . What does FDA say regarding BLA/NDA stage assays? The accuracy, sensitivity, specificity, and reproducibility of test methods employed by the firm shall be established and documented.

3 Such validation and documentation may be accomplished in accordance with . 21 CFR (e) (Testing and release for distribution). At the time the application is submitted to the regulatory authorities, applicants should have validated the analytical procedures uses in the specifications. ICH Q6B. Analytical methods should be validated . ICH Q7. The stability program shall include reliable, meaningful, and specific test methods. 5. 21 CFR (a)(3). What does FDA say regarding IND stage assays? Although in each phase of the investigation sufficient information is required to assure the proper identification, quality, purity, and strength of the investigational drug, the amount of information needed to make that assurance will vary with each phase of the investigation, the proposed duration of the investigation, the dosage form, and the amount of information otherwise available.

4 21 CFR (a)(7)(i). The {IND} submission is required to contain: the acceptable limits and analytical methods used to assure the identity, strength, quality, and purity of the drug substance [drug product] . 21 CFR (a)(7)(iv)(a) [21 CFR (a)(7)(iv)(b)]. 6. One of the clearest statements FDA makes regarding a requirement for Assay validation at the IND stage is in the PROCESS validation guidance? While validated analytical methods are not required during product- and process-development activities, methods should be scientifically sound ( , specific, sensitive, and accurate), suitable, and reliable for the specified purpose. Guidance for Industry: Process validation : General Principles and Practices. FDA/CDER, CBER, CVM. 2008. 7. validation vs. qualification Are we speaking the same language? Process validation is defined as the collection and evaluation of data, from the process design stage throughout production, which establishes scientific evidence that a process is capable of consistently delivering quality products.

5 Process validation involves a series of activities taking place over the lifecycle of the product and process. This guidance describes the process validation activities in three stages. Stage 1 Process Design: The commercial process is defined during this stage based on knowledge gained through development and scale-up activities. Stage 2 Process qualification : During this stage, the process design is confirmed as being capable of reproducible commercial manufacturing. Stage 3 Continued Process Verification: Ongoing assurance is gained during routine production that the process remains in a state of control. Guidance for Industry: Process validation : General Principles and Practices. FDA/CDER, CBER, CVM. 2008. 8. validation vs. qualification Are we speaking the same language? Lifecycle Stage 1 Process Design: The commercial process is defined during this stage based on knowledge gained through development and scale-up activities.

6 Stage 2 Process qualification : During this stage, the process design is confirmed as being capable of reproducible commercial manufacturing. Stage 3 Continued Process Verification: Ongoing assurance is gained during routine production that the process remains in a state of control. 9. validation vs. qualification Are we speaking the same language? Lifecycle (borrowing from Process validation ). Stage 1 Assay Design: The Assay is defined during this stage based on knowledge gained through development activities. Stage 2 Assay qualification : During this stage, the Assay design is confirmed as being capable of producing reproducible results suitable for the specified purpose. Stage 3 Continued Assay Verification: Ongoing assurance is gained during routine use that the Assay remains in a state of control. 10. validation vs. qualification Are we speaking the same language?

7 For a bioanalytical [BA, BE, PK] method to be considered valid, specific acceptance criteria should be set in advance and achieved for accuracy and precision for the validation of QC samples over the range of the standards. The validity of an analytical method should be established and verified by laboratory studies, and documentation of successful completion of such studies should be provided in the Assay validation report. Guidance for Industry: Bioanalytical Method validation . FDA/CDER, CVM. 2001. 11. validation vs. qualification Are we speaking the same language? qualification : The Assay design is confirmed as being capable of producing reproducible results suitable for the specified purpose. Test how the Assay performs and decide if that performance is suitable for that point in development. If it isn't good enough, change the Assay . Work in progress, but scientifically sound and suitable for its purpose.

8 validation : The Assay is tested against specific acceptance criteria set in advance to verify that the performance characteristics (those listed in ICH Q2(R1) and potentially others, depending on the Assay and the product) of the final method are suitable and reliable for the intended applications. Documentation of successful completion of such studies should be provided in an Assay validation report. Final product, documented to meet performance acceptance 12. criteria. validation vs. qualification Are we speaking the same language? Lifecycle (borrowing from Process validation ). Stage 1 Assay Design: The Assay is defined during this stage based on knowledge gained through development activities. Stage 2 Assay qualification : During this stage, the Assay design is confirmed as being capable of producing reproducible results suitable for the specified purpose. Scientifically sound work in progress.

9 Stage 2 - Formal Assay validation Study: The Assay is tested against specific acceptance criteria set to verify that the performance characteristics of the final method are suitable and reliable for the intended applications. Stage 3 Continued Assay Verification: Ongoing assurance is gained during routine production that the Assay remains in a state of control. (system suitability, Assay suitability, trending of results, OOS, etc.) 13. Early Development Submissions need to contain enough information for reviewers to determine whether or not the methods are scientifically sound ( , specific, sensitive, and accurate), suitable, and reliable for the specified purpose. The specified purpose is to provide sufficient information to assure the proper identification, quality, purity, and strength. The Assay needs to be sufficiently developed to ensure that acceptance criteria are meaningful.

10 What does this really mean? 14. Early Development Reviewers need enough information to make these assessments. How much information is needed will be product and Assay dependent. concentration potency less CE methods more UV spectroscopy cell based Assay 15. Early Development Reviewers need enough information to make these assessments. How much information is needed will be product and Assay dependent. cIEF cIEF cIEF. less more Charge variants Antibody drug conjugate of an IgG1 Charge variants drug:antibody ratio of a Fc fusion protein 16. Early Development Reviewers need enough information to make these assessments. How much information is needed will be product and Assay dependent. cIEF for identity example: If the product is a second generation product, a sponsor may need to include some specificity data in the IND original submission. The data themselves can sometimes provide information regarding the suitability/reliability of the Assay .