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.
Choi
Dong Hwan
DH
Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.
Katakura
Yoshio
Y
Matsuda
Rieko
R
Hayashi
Yuzuru
Y
Ninomiya
Kazuaki
K
Shioya
Suteaki
S
D000465:Algorithms; D000906:Antibodies / Q000737:chemistry*; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000737:chemistry*; D003198:Computer Simulation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004798:Enzymes / Q000737:chemistry*; D008956:Models, Chemical; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000906:Antibodies; D000941:Antigens; D004798:Enzymes