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Challenges in selectivity, specificity and quantitation range of ligand-binding assays: case studies using a
Tong-Yuan Yang1, David J Uhlinger, Stephen A Ayers
1Biologics Clinical Pharmacology, Biotechnology Center of Excellence, Janssen R&D, LLC, Spring House, PA 19477, USA.
Bioanalysis
|May 17, 2014
Summary
Microfluidic immunoassay systems offer improved ligand-binding assay (LBA) performance over traditional plate-based methods. These systems reduce background noise and matrix interference, enhancing selectivity and quantitation range for bioanalysis.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Assay Development
Background:
- Plate-based ligand-binding assays (LBAs) present challenges in achieving desired selectivity, specificity, and quantitation range.
- Method development for LBAs often encounters issues like nonspecific background signal and matrix interference.
Purpose of the Study:
- To compare the performance of plate-based LBAs with microfluidic immunoassay systems.
- To evaluate different immunoassay platforms for method development in pharmacokinetic and pharmacodynamic studies.
Main Methods:
- Comparative case studies of plate-based and microfluidic ligand-binding immunoassay platforms.
- Method development focused on selectivity, specificity, and range of quantitation.
Main Results:
- Plate-based LBAs struggle with nonspecific background, matrix effects, linearity, and drug interference.
- Microfluidic systems demonstrate faster kinetics, reducing background and matrix effects.
- Microfluidic assays showed increased assay linear range and improved drug tolerance.
Conclusions:
- Microfluidic immunoassay systems offer superior performance for LBAs affected by matrix effects.
- Short incubation times in microfluidics benefit LBAs, outperforming traditional platforms like Meso Scale Discovery and ELISA in bioanalysis.
