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Toxins 2017, 9(12), 386; https://doi.org/10.3390/toxins9120386

Detection of Abrin Holotoxin Using Novel Monoclonal Antibodies

Western Regional Research Center, U.S. Department of Agriculture, Agricultural Research Service, 800 Buchanan Street, Albany, CA 94710, USA
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Academic Editors: Tomas Girbes and Nilgun E. Tumer
Received: 8 September 2017 / Revised: 9 November 2017 / Accepted: 23 November 2017 / Published: 28 November 2017
(This article belongs to the Section Plant Toxins)
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Abstract

Abrin, a member of the ribosome-inactivating protein family, is produced by the Abrus precatorius plant. Having the potential to pose a severe threat to both human and animal health, abrin is classified as a Select Agent by the U.S. Department of Health and Human Services. However, an immunoassay that is specific for intact abrin holotoxin has not yet been reported. In this study, seven new monoclonal antibodies (mAbs), designated as Abrin-1 through Abrin-7 have been developed. Isotyping analyses indicate these mAbs have IgG1, IgG2a, or IgG2b heavy-chains and kappa light-chains. Western blot analyses identified two abrin A-chain specific mAbs, Abrin-1 and Abrin-2, and four B-chain specific mAbs (Abrin-3, -5, -6, and -7). A sandwich enzyme-linked immunosorbent assay (ELISA), capable of detecting a mixture of abrin isoforms and agglutinins was developed using B-chain specific Abrin-3 for capture and A-chain specific Abrin-2 as detector. The ELISA is highly sensitive and detects 1 ng/mL of the abrin holotoxin in phosphate-buffered saline, nonfat milk, and whole milk, significantly below concentrations that would pose a health concern for consumers. This ELISA also detects native abrin in plant extracts with a very low background signal. The new abrin mAbs and ELISA should be useful for detecting this potent toxin in the milk supply chain and other complex matrices. View Full-Text
Keywords: abrin; agglutinin; enzyme-linked immunosorbent assay; milk; monoclonal antibody; ricin; Western blot abrin; agglutinin; enzyme-linked immunosorbent assay; milk; monoclonal antibody; ricin; Western blot
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He, X.; Patfield, S.; Cheng, L.W.; Stanker, L.H.; Rasooly, R.; McKeon, T.A.; Zhang, Y.; Brandon, D.L. Detection of Abrin Holotoxin Using Novel Monoclonal Antibodies. Toxins 2017, 9, 386.

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