Challenges in selectivity, specificity and quantitation range of ligand-binding assays: case studies using a microfluidics platform
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Published on: December 10, 2011
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.
Background:
Method developers of plate-based ligand-binding assays (LBAs) often face challenges establishing selectivity, specificity and range of quantitation to meet the needs of a particular study. Case studies are presented to compare different ligand-binding immunoassay platforms (plate-based vs microfluidic system) in method development to support pharmacokinetic and pharmacodynamic studies.
Results:
Studies highlight the challenges of plate-based LBAs to establish selectivity, specificity and range of quantitation as a result of nonspecific background signal, matrix interference, lack of linearity and drug interference. The fast assay kinetics of a microfluidic immunoassay system was shown to generally reduce nonspecific background and matrix effects, while increasing assay linear range and drug tolerance.
Conclusion:
The short incubation times with microfluidics can be beneficial for LBAs burdened by matrix effects and in these cases had superior assay performance compared with widely used immunoassay platforms in bioanalysis, for example, Meso Scale Discovery(®) and enzyme-linked immunosorbent assay.
