Development of a homogeneous immunoassay system using protein A fusion fragmented Renillaluciferase

Abstract

A homogeneous immunoassay system was developed using fragmented Renillaluciferase (Rluc). The B domain of protein A was fused to two Rluc fragments. When complexes between an antibody and fragmented Rluc fusion proteins bind to target molecules, the Rluc fragments come into close proximity and the luminescence activity of fragmented Rluc is restored by complementation. As proof-of-principle, this fragmented Rluc system was used to detect E. coli homogeneously using an anti-E. coliantibody.