A two site enzyme‐linked immunosorbent assay for wheat gliadins has been developed. Polyclonal antibodies from chicken eggs were used as the capture agent adsorbed to the surface of the microtitration plate. Murine monoclonal antibodies bound to captured gliadin were detected by addition of enzyme‐labelled, species‐specific antibodies. The assay was developed for the purposes of screening monoclonal antibodies specific for conformational epitopes on gliadins. The properties of a panel of 6 anti‐gliadin antibodies in the assay were assessed and compared to properties exhibited under conditions where conformational epitopes were less likely to be present. Three of the monoclonal antibodies appeared to be specific for conformational epitopes.