Least detectable concentration and dynamic range of three immunoassay systems using the same antibody.
The least detectable concentration (LDC) and dynamic range (DR) of three immunoassay systems are compared using four distinct antibodies (all specific for estradiol but with different affinities) on each system. The systems evaluated include the industry standard, ELISA, and two biosensors, surface plasmon resonance and kinetic exclusion. In all cases, the measurements of inhibition curves (response vs estradiol concentration) were contracted to outside experts (the biosensor manufacturers themselves in the case of the biosensors), each of whom was supplied with the same blind samples. Each biosensor manufacturer also reported an estimate of the equilibrium dissociation constant (Kd) for each of the antibodies. The LDC and DR observed for the kinetic exclusion biosensor are consistent with an interpretation of Kd limited detection while that from the other biosensor and ELISA show limits of detection somewhat above those expected for Kd limited performance. The determined LDC and DR of each biosensor is self-consistent in the sense that none of the inhibition data contradicts theoretical limits associated with the Kd as measured on that system; however, some contradictions are apparent across platforms. The use of multiple antibodies of differing Kd improves confidence that the observed differences in performance are associated with the immunoassay system rather than the particular analyte.
Glass
Thomas R
TR
Central Research Institute of the Electric Power Industry, Department of Bioscience, 1646 Abiko, Abiko City, Chiba, Japan.
Ohmura
Naoya
N
Saiki
Hiroshi
H
D000911:Antibodies, Monoclonal / Q000276:immunology*; D015374:Biosensing Techniques / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000032:analysis* / Q000276:immunology; D007118:Immunoassay / Q000379:methods*; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D004958:Estradiol