Related Experiment Video
Updated: May 17, 2026
08:44
Dried Blood and Serum Spots As A Useful Tool for Sample Storage to Evaluate Cancer Biomarkers
Published on: June 11, 2018
Validation of a ligand binding assay using dried blood spot sampling
Daniel Burns1, Laura Brunner, Surendran Rajendran
1Pharmacokinetics and Drug Metabolism Department, Amgen Inc., Mail Stop 30E-3-C, One Amgen Center Drive, Thousand Oaks, CA 91320, USA.
The AAPS Journal
|November 9, 2012
Summary
Dried blood spots (DBS) offer a microsampling alternative for drug development. This study validates a ligand binding assay for a monoclonal antibody using DBS, establishing a model for future large molecule therapeutic analysis.
Area of Science:
- Pharmacokinetics and Drug Development
- Biomarker Analysis
- Analytical Chemistry
Background:
- Dried blood spots (DBS) are an established microsampling technique for small molecule drug development.
- The application of DBS for large molecule therapeutics, such as monoclonal antibodies, is not yet well-established.
- Ligand binding assays (LBAs) are crucial for quantifying large molecule therapeutics.
Purpose of the Study:
- To describe the method validation of a ligand binding assay (LBA) for dried blood spot (DBS) sampling of a therapeutic monoclonal antibody (AMG 162/Denosumab).
- To evaluate the suitability of DBS for large molecule therapeutic monitoring.
- To establish a model process for future LBA method validation using DBS for large molecules.
Main Methods:
- Modification of an existing serum LBA for DBS analysis.
- Fit-for-purpose method validation including accuracy, precision, selectivity, dilutional linearity, and stability.
- Evaluation of DBS-specific parameters: spot volume, extraction recovery, whole blood stability, and hematocrit effects.
Main Results:
- The validated LBA demonstrated assay robustness with inter-assay precision ≤ 19% and accuracy ≤ 9%.
- Selectivity, extraction recovery, dilutional linearity, and stability were successfully demonstrated.
- Potential limitations identified include hematocrit effects and the use of whole blood for standards/quality controls.
Conclusions:
- This study presents the first thorough method validation of an LBA using DBS for a therapeutic monoclonal antibody.
- The validated method is suitable for quantifying therapeutic monoclonal antibodies in DBS samples.
- The established process serves as a model for validating LBAs for other large molecule therapeutics or biomarkers using DBS.
