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A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin

Author:
Christina C. Tam, Luisa W. Cheng, Xiaohua He, Paul Merrill, David Hodge, Larry H. Stanker
Source:
Toxins 2017 v.9 no.10 pp. e328
ISSN:
2072-6651
Subject:
Abrus precatorius, Western blotting, abrin, agglutinins, antibody detection, detection limit, enzyme-linked immunosorbent assay, glycosidases, monoclonal antibodies, polyclonal antibodies, rapid methods, saponins, toxicity
Abstract:
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.
Agid:
6472266
Handle:
10113/6472266