Challenges in selectivity, specificity and quantitation range of ligand-binding assays: case studies using a microfluidics platform.
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.


CONCLUSIONS
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.
Biologics Clinical Pharmacology, Biotechnology Center of Excellence, Janssen R&D, LLC, Spring House, PA 19477, USA.
Tong-Yuan
TY
Yang
David J
DJ
Uhlinger
Stephen A
SA
Ayers
Denise M
DM
O'Hara
Alison P
AP
Joyce
D015415:Biomarkers / Q000032:analysis; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods*; D008024:Ligands; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D012680:Sensitivity and Specificity
D015415:Biomarkers; D008024:Ligands