Simulation model for predicting limit of detection and range of quantitation of competitive enzyme-linked immunosorbent assay
The limit of detection (LOD) and range of quantitation (ROQ) of competitive enzyme-linked immunosorbent assay (ELISA) were determined from a model describing the calibration curve and precision profile of the assay. The calibration curve is given by solving the differential equations describing the change in the concentrations of an antigen–antibody complex and an enzyme-conjugated antigen–antibody complex by a Runge–Kutta method. The precision profile is described in terms of possible error sources such as the pipetting volumes of the analyte, enzyme-conjugated antigen, antibody and substrate solutions, calibration curve and inherent absorbances between the wells in an ELISA plate. An appropriate concentration of the enzyme-conjugated antigen that balances a low detection limit and sufficient color development was found to be in a narrow range, which is consistent with the empirical rule. The optimum conditions for competitive ELISA using an antibody with a kinetic property can be designed from our model.
Journal: Journal of Bioscience and Bioengineering - Volume 103, Issue 5, May 2007, Pages 427–431