Analysis of Castor by ELISAs that Distinguish Ricin and Ricinus communis agglutinin (RCA)

To facilitate the analysis of castor (Ricinus communis L.) seed fractions and germplasm for ricin content, we investigated the use of enzyme-linked immunosorbent assay (ELISA) methods to differentiate between ricin toxin and the related Ricinus communis agglutinin (RCA). Both proteins are based on a heterodimeric AB structure, with a common A chain. RCA comprises two dimers of A and B chains. Both proteins are found in the meal remaining after castor oil extraction and impede the commercial production of castor seed in the USA. We identified pairs of monoclonal antibodies (mAbs) that could distinguish between the structurally related proteins that share a common A chain. Antibody specificity was determined by ELISA and checked by immunoblotting. We found that mAb–mAb pairs afforded quantification of each castor protein, and that a mAb paired with a commercial polyclonal antibody provided detection of both with comparable sensitivity.