Development and use of a monoclonal antibody to detect semi-digested proteins of the English grain aphid,Sitobion avenae, in the guts of ladybird beetle predators

A monoclonal antibody (McAb), EGA-4A9, was developed to detect the semi-digested proteins of the English grain aphid, Sitobion avenae (Fabricius) (Hemiptera: Aphididae), in predatory ladybird beetles (species of the genera Adonia, Coccinella, Hippodamia, and Propylea) using the gut homogenate of Adonia variegata (Goeze) (Coleoptera: Coccinellidae) adults which had fed on S. avenae as immunogen. The McAb was selected by screening hybridoma lines for antibodies that bound with the semi-digested aphid proteins in ladybirds. A double antibody sandwich enzyme-linked immuno-sorbent assay (ELISA) using ClonotypingTM System/HRP showed that it belonged to the IgG2a isotype. It did not cross-react with any of the 21 arthropod species tested besides the ladybird beetles fed on S. avenae with an indirect ELISA. It could still detect the semi-digested proteins in the gut of a ladybird adult, kept at 25 °C, that had ingested one aphid 6 days before. The extended antigen detection period and the high specificity of the antibody indicated that EGA-4A9 could be used to study interactions between English grain aphids and their ladybird predators in the field. Between 28 and 72% of coccinellids collected from the field were positive for English grain aphid protein by ELISA. The percentage of McAb-positive predatory ladybird beetles was positively correlated with the density of S. avenae in wheat fields