Immunoassay principle based on exchange reactions: investigations on agarose gel and silanized quartz

Abstract

Preliminary studies of a new immunoassay principle based on exchange reactions is reported. Exchange of 125I-labelled insulin with unlabelled insulin from immobilized monoclonal antibodies was investigated. From antibody immobilized on a gel substrate the tagged insulin was exchanged according to a first-order process. With antibody immobilized on a quartz substrate by two different methods, the kinetics was changed dramatically, probably because of the non-specific interaction between the ligand and the surface. The recorded adsorption isotherms could not be described by the Langmuir adsorption equation, and a model allowing for non-specific adsorption of the ligand was developed. This model gave a satisfactory fit to the experimental data, allowing computation of adsorption parameters. It is concluded that even the best method used to immobilize receptors on quartz is not adequate for an exchange assay to be made. However, this coating method may lead to more sensitive receptor-based assays of more conventional type.