Flow Injection Renewable Surface Immunoassay for Real Time Monitoring of Biospecific Interactions

This is the authors' abstract. We don't add key points for this paper.

An automated system for performing rapid immunoassay, kinetic measurements, and affinity ranking of biomolecular interactions using fluorescence-labeled ligands is described. Its distinctive feature is the automated renewal of solid phase for each measurement, which avoids the need for regeneration of the sensing surface. This system--flow injection renewable surface immunoassay (FIRSI)--is used for the first time here for determination of rate constants for an antibody/antigen interaction and for affinity ranking of several related antigens against one antibody. The performance of the system is compared with a commercial BIAcore system that uses surface plasmon resonance for monitoring biomolecular interactions. While the values of association and dissociation rate constants for human serum albumin (HSA) with monoclonal anti-HSA antibody obtained by these techniques were comparable, it is shown that the FIRSI techniques requires simpler instrumentation, handles a broader size range of analytes, and does not suffer from disturbances caused by changes in the refractive index.