Monoclonal antibody-based enzyme immunoassay for detection of ricin |
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Authors: | Shyu Huey-Fen Chiao Der-Jiang Liu Hwan-Wun Tang Shiao-Shek |
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Affiliation: | Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, Republic of China. |
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Abstract: | A sensitive and specific enzyme-linked immunoadsorbent assay (ELISA) was developed to detect ricin in biological fluids. The assay is based on the sandwich format using monoclonal antibodies (MAbs) of two distinct specificities. An affinity-purified anti-ricin B chain MAb (1G7) is utilized to adsorb ricin from solution and the second anti-ricin A chain MAb (5E11) conjugated with peroxidase is then used to form a sandwich, and peroxidase allows color development and measurement of optical density at 450 nm. Standard curves were linear over the range of 2.5-100 ng/mL ricin. The limit of detection was below 5 ng/mL in assay buffer as well as in a 1:10 dilution of urine or 1:50 dilution of human serum spiked with ricin. |
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