Adjusting factors such as pH, temperature, incubation time, and concentrations of reagents can all minimize cross-reactivity.
Cross-reactivity decreases as the incubation time increases, reaching a minimum when the reaction reaches equilibrium. Higher temperatures speed up reaction rates, helping to reach equilibrium faster. Thus, raising the incubation temperature can reduce cross-reactivity by promoting quicker equilibrium.
Using assays with sensitive detection and lower concentrations of reagents reduces the likelihood of cross-reactivity, leading to more accurate and specific results. Switching to lower concentrations of reagents reduces cross-reactivities by as much as five times.
Additionally, using multiple antibodies that target different epitopes of the antigen can decrease the risk of cross-reactivity. Since each antibody binds to a distinct epitope, they are less likely to bind to non-specific targets, thus reducing the chances of false signals.