Comparison of glycosphingolipids and antibodies as receptor molecules for ricin detection.
ABSTRACT Glycosphingolipids (GSLs) have been shown to undergo strong interactions with a number of protein toxins, including potential bioterrorism agents such as ricin and botulinum neurotoxin. Characterization of this interaction in recent years has led to a number of studies where GSLs were used as the recognition molecules for biosensing applications. Here, we offer a comparison of quartz crystal microbalance (QCM) sensors for the detection of ricin using antibodies and the GSLs GM1 and asialoGM1, which have been shown to undergo strong interactions with ricin. The presence, orientation, and activity of the GSL and antibody films were confirmed using ellipsometry, Fourier transform infrared spectroscopy (FT-IR), and QCM. It was found that the GSLs offered more sensitive detection limits when directly compared with antibodies. Both GSLs had lower detection limits at 5 microg/mL, approximately 5 times lower than were found for antibodies (25 microg/mL), and their linear detection range extended to the highest concentrations tested (100 microg/mL), almost an order of magnitude beyond the saturation point for the antibody sensors. Potential sites for nonspecific adsorption were blocked using serum albumin without sacrificing toxin specificity.
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ABSTRACT: Ricin is a plant toxin with high bioterrorism potential due to its natural abundance and potency in inducing cell death. Early detection of the active toxin is essential for developing appropriate countermeasures. Here we review concepts for designing ricin detection methods, including mechanism of action of the toxin, advantages and disadvantages of current detection assays, and perspectives on the future development of rapid and reliable methods for detecting ricin in environmental samples. Copyright © 2014. Published by Elsevier Inc.Biotechnology Advances 12/2014; 33(1). · 8.91 Impact Factor
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ABSTRACT: Ricin is a highly toxic, dichain ribosome-inactivating protein present in the seeds of Ricinus communis (castor), grown principally as a source of high quality industrial lubricant and as an ornamental. Because of its presence in industrial byproducts and its documented use for intentional poisoning, there is a need for analytical methodology to quantify ricin in both castor extracts and food matrices. We developed a panel of monoclonal antibodies to ricin, with most having strong cross-reactivity with RCA-1, a homologous but less toxic castor agglutinin. Some of the IgM-producing hybridomas appeared to produce a second, IgG isotype and were further analysed by fluorescence-activated cell sorting. The antibodies were effective in various ELISA formats, many with IC50's in the range of 0.1–10 ng/mL and minimal matrix effects in skim milk. Assay specificity can be adjusted for analytical needs by varying the combination of antibodies in a sandwich ELISA format.Food and Agricultural Immunology 03/2009; 20(1):11-22. · 0.98 Impact Factor
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ABSTRACT: Biological agents including viruses, bacteria, and other naturally occurring pathogenic organisms, along with the toxins they produce, are considered far harder to detect and defend against than chemical agents. Here we provide an overview of the predominant molecular sensing technologies for the detection of these agents. This includes biosensing strategies based upon use of antibodies, genomic analysis, biochemical testing, other recognition interactions, and cellular-based responses. We survey some popular sensing approaches, illustrate them with current examples showing how they have been applied, and discuss their intrinsic benefits and potential liabilities. Lastly, within the context of security applications, some approaches for integrating sensing technologies into field-portable devices are discussed.Materials Today 03/2008; 11(3):38-49. · 10.85 Impact Factor