Biomarker Assay Validation by Mass Spectrometry.
Carmen Fernández-Metzler1, Brad Ackermann2, Fabio Garofolo3
1PharmaCadence Analytical Services, LLC, 1440 Industry Road, Hatfield, PA, 19940, USA. carmen.fernandez-metzler@PharmaCadence.com.
The AAPS Journal
|May 9, 2022
Summary
This study provides recommendations for biomarker assay validation (BAV) using LC-MS, emphasizing fit-for-purpose (FFP) validation tailored to the context of use (COU). These guidelines ensure reliable biomarker data for regulatory submissions.
Area of Science:
- Analytical Chemistry
- Biomarker Discovery and Validation
- Regulatory Science
Background:
- Pharmacokinetic (PK) and toxicokinetic (TK) assay validation is established, with FDA guidance available.
- Biomarker assay validation presents unique challenges, including analyte presence in matrices and reference standard procurement.
- Existing guidance often focuses on challenges rather than specific experimental designs for biomarker assay validation.
Purpose of the Study:
- To provide specific recommendations for biomarker assay validation (BAV) using Liquid Chromatography-Mass Spectrometry (LC-MS).
- To address the unique challenges of validating assays for both small and large molecule biomarkers.
- To promote a fit-for-purpose (FFP) approach to biomarker assay validation, aligning with the context of use (COU).
Main Methods:
- Review and synthesis of existing guidance on bioanalytical method validation (BMV).
- Adaptation of PK assay validation principles to biomarker applications.
- Development of consensus recommendations for biomarker assay validation plan and acceptance criteria.
Main Results:
- Proposed a framework for biomarker assay validation (BAV) emphasizing the context of use (COU).
- Recommended integrating relevant PK assay validation elements into biomarker assay validation.
- Defined specific validation elements and acceptance criteria tailored for biomarker assays, including LC-MS.
Conclusions:
- Biomarker assay validation requires a fit-for-purpose (FFP) strategy, dictated by the assay's context of use (COU).
- A comprehensive validation plan, including COU definition and adapted validation elements, is crucial for reliable biomarker data.
- These recommendations facilitate robust biomarker assay validation for regulatory submissions.
More Related Videos
13:00Low Molecular Weight Protein Enrichment on Mesoporous Silica Thin Films for Biomarker Discovery
Published on: April 17, 2012
13.6K
11:09An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry
Published on: April 17, 2018
10.3K
Related Concept Videos
01:19MALDI-TOF Mass Spectrometry
5.4K
Mass spectrometry is a powerful characterization technique that can identify and separate a wide variety of compounds ranging from chemical to biological entities, based on their mass-to-charge ratio (m/z). The instruments that allow this detection, known as mass spectrometers, have three components: an ion source, a mass analyzer, and a detector. These spectrometers differ based on the nature of their ion source and analyzers.
Matrix-assisted laser desorption ionization (MALDI) is a commonly...
Matrix-assisted laser desorption ionization (MALDI) is a commonly...
5.4K
01:33Peptide Identification Using Tandem Mass Spectrometry
6.9K
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
6.9K
01:21Mass Spectrometry: Complex Analysis
914
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
914
01:19Mass Spectrometry: Overview
6.2K
Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electrospray ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave...
6.2K
01:08Matrix-Assisted Laser Desorption Ionization (MALDI)
466
Matrix-assisted laser desorption ionization (MALDI) is a powerful analytical technique used in mass spectrometry. It enables the identification and characterization of various biomolecules, including proteins, peptides, nucleic acids, and carbohydrates. MALDI spectrometry is widely employed in biological and medical research, as well as in fields like pharmacology and biochemistry.
The analyte of interest, a biomolecule or a mixture of biomolecules, is mixed with a suitable matrix material. The...
The analyte of interest, a biomolecule or a mixture of biomolecules, is mixed with a suitable matrix material. The...
466
01:16Mass Spectrometers
6.0K
This lesson details the instrumentation of a mass spectrometer—a physical instrument to perform mass spectrometry on analyte molecules and record the characteristic mass spectra. This is achieved via three chief functions:
6.0K