On-line flow displacement immunoassay for fatty acid-binding protein

In standard displacement flow immunoassays the analyte in the sample creates an active dissociation of labelled antigens (or antigen homologues) from an antigen binding site of an immobilized antibody, after which the labelled substance is measured downstream. Such systems have been described for molecules up to 1 kDa. In this study, we demonstrate displacement in a flow system for the detection of a small protein, cytoplasmic heart-type fatty acid-binding protein (15 kDa), a plasma marker for myocardial injury. The displacement system uses an inverse set-up: enzyme labelled monoclonal antibodies are associated to immobilized antigen, and are displaced by analyte in the sample. The system permits detection of both physiological (2- 12 μg l^-1) and pathological concentrations (12-2000 μg l^-1) of fatty acid-binding protein in an on-line flow system.