Modelling random antibody adsorption and immunoassay activity

One of the primary considerations in immunoassay design is optimizing the concentration of capture antibody in order to achieve maximal antigen binding and, subsequently, improved sensitivity and limit of detection. Many immunoassay technologies involve immobilization of the antibody to solid surfaces. Antibodies are large molecules in which the position and accessibility of the antigen-binding site depend on their orientation and packing density. In this paper we propose a simple mathematical model, based on the theory known as random sequential adsorption (RSA), in order to calculate how the concentration of correctly oriented antibodies (active site exposed for subsequent reactions) evolves during the deposition process. It has been suggested by experimental studies that high concentrations will decrease assay performance, due to molecule denaturation and obstruction of active binding sites. However, crowding of antibodies can also have the opposite effect by favouring upright orientations. A specific aim of our model is to predict which of these competing effects prevails under different experimental conditions and study the existence of an optimal coverage, which yields the maximum expected concentration of active particles (and hence the highest signal).