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A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin

Christina C. Tam, Luisa W. Cheng, Xiaohua He, Paul Merrill, David Hodge, Larry H. Stanker
Toxins 2017 v.9 no.10 pp. e328
Abrus precatorius, Western blotting, abrin, agglutinins, antibody detection, detection limit, enzyme-linked immunosorbent assay, glycosidases, monoclonal antibodies, polyclonal antibodies, rapid methods, saponins, toxicity
Abrin, one of the most highly potent toxins in the world, is derived from the plant, Abrus precatorius. Because of its high toxicity, it poses potential bioterror risks. Therefore, a need exists for new reagents and technologies that would be able to rapidly detect abrin contamination as well as lead to new therapeutics. We report here a group of abrin-specific monoclonal antibodies (mAbs) that recognize abrin A-chain, intact A-B chain toxin, and agglutinin by Western blot. Additionally, these mAbs were evaluated for their ability to serve as capture antibodies for a sandwich (capture) ELISA. All possible capture-detector pairs were evaluated and the best antibody pair identified and optimized for a capture ELISA. The capture ELISA based on this capture-detector mAb pair had a limit of detection (L.O.D) of ≈1 ng/mL measured using three independent experiments. The assay did not reveal any false positives with extracts containing other potential ribosome-inactivating proteins (RIPs). Thus, this new capture ELISA uses mAbs for both capture and detection; has no cross-reactivity against other plant RIPs; and has a sensitivity comparable to other reported capture ELISAs using polyclonal antibodies as either capture or detector.