A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin
- 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
- https://doi.org/10.3390/toxins9100328