An integrated model for bead-based immunoassays

Abstract

Bead-based immunoassays have shown great promise for rapid and sensitive protein quantification. However, there still lacks holistic understanding of assay performance that can inform assay design and optimization. In this paper, we present an integrated mathematical model for surface coverage bead-based assays. This model examines the building blocks of surface coverage assays, including heterogeneous binding of analyte molecules on bead or sensor surfaces, attachment of bead labels to sensor surfaces, and generation of electrochemical current by bead labels. To demonstrate and validate this model, we analyze a semi-homogeneous bead-based electronic enzyme-linked immunosorbent assay and find that experimental results agree with various model predictions. We show that the model can provide design guidance for choice of various assay parameters including bead size, bead number, antibody affinity and assay time, and provide a perspective to reconcile the performance of various implementations of surface coverage assays.