Bioanalytical Method Validation for Biomarkers Guidance

FDA has published a guidance titled "Bioanalytical Method Validation for Biomarkers” to help sponsors of INDs and applicants of NDAs, BLAs, sNDAs, sBLAs, and ANDAs to validate bioanalytical methods used to evaluate biomarker concentrations. The document states that it can also inform the development of bioanalytical methods for analyzing biomarker concentrations in nonclinical study samples.

“The recommendations in this guidance pertain only to the validation of bioanalytical assays to measure in vivo biomarker concentrations in biological matrices such as blood or urine,” FDA says. “This guidance does not apply to bioanalytical method validations for the measurement of veterinary drug concentrations or veterinary biomarker concentrations.”

FDA says that biomarkers can be used for a wide variety of purposes during drug development. “Therefore, a fit-for-purpose approach should be used when determining the appropriate extent of method validation,” it says. For example, when biomarker data will be used to support regulatory decision-making, such as the pivotal determination of safety and/or effectiveness to support approval or to support dosage instructions in product labeling, the assay should be fully validated as described in the guidance for industry M10 Bioanalytical Method Validation and Study Sample Analysis (November 2022). Alternatively, for assays intended to only support internal pharmaceutical company decision-making (e.g., candidate selection, go-no-go decisions, proof-of-concept), the sponsor should incorporate the extent of method validation they deem appropriate.”

FDA says that validating an analytical method ensures that the data are reliable by addressing certain key questions, including:

• Does the method measure the intended analyte? For example, does anything interfere with the measurement, and is the method specific or selective for the analyte?
• What is the variability associated with these measurements? For example, what are the accuracy and precision of the method?
• What is the range in measurements that provides reliable data? For example, what is the sensitivity of the method (e.g., what is the lower limit of quantitation of the method and upper limit of quantitation of the method?
• How do sample collection, handling, and storage affect the reliability of the data from the bioanalytical method? For example, what steps need to be followed while collecting samples? Do the samples need to be frozen during shipping? What temperatures are required to store the samples, and how long can the samples be stored? How are samples affected by benchtop manipulations?

Additionally, the agency notes that method validation for biomarker assays should address the same questions as method validation for drug assays. “The accuracy, precision, sensitivity, selectivity, parallelism, range, reproducibility, and stability of a biomarker assay are important characteristics that define the method,” it says. “The approach described in the guidance for industry M10 Bioanalytical Method Validation and Study Sample Analysis (November 2022) for drug assays should be the starting point for validation of biomarker assays, especially chromatography and ligand-binding-based assays.”