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Use of a Heterologous Solid-Phase Antigen in an Indirect Competitive Antibody-Capture Enzyme-Linked Immunosorbent Assay for T-2 Mycotoxin

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Abstract:

Anti-T-2 toxin (T-2) antibodies raised in chicken hosts were isolated as total IgY (chicken immunoglobulins of IgY isotype) from serum and the egg yolk. They were used for quantitation of T-2 toxin in an indirect competitive enzyme-linked immunosorbent assay (ELISA) which employed molecularly distinct toxin-carrier conjugates as solid-phase assay antigens. The sensitivities of the assays utilizing any of the purified IgY or unfractionated serum were similar but varied with respect to the type of conjugate used and could be improved by lowering the concentration of assay antigens or antibodies. Low concentrations of both immunoreactants resulted in assays of superb sensitivity, although the standard curves generated with some assay antigens decreased markedly in slopes and impractically long incubation times were required to develop sufficiently high read-out signals. However, high optical signals were obtained in a very short period of time when an amplified substrate system instead of para-nitrophenyl phosphate (pNPP) was used for the detection of a reporter enzyme, alkaline phosphatase. The use of an amplified substrate and of a solid-phase antigen that was prepared with Iso T-2 toxin attached directly to a carrier protein yielded an ELISA that had an improved ability to detect T-2.

Keywords: AFFINITY CHROMATOGRAPHY; BRIDGE AND HAPTEN HETEROLOGY; ELISA AMPLIFICATION; IGY ANTIBODIES; T-2 MYCOTOXIN

Document Type: Research Article

Affiliations: 1: Department of Animal Science, Faculty of Food and Agricultural Sciences, University of Manitoba, Winnipeg, MB, Canada R3T 2N2; Cangene Corporation, 104 Chancellor Matheson Road, Winnipeg, MB, Canada R3T 5Y3 2: Department of Animal Science, Faculty of Food and Agricultural Sciences, University of Manitoba, Winnipeg, MB, Canada R3T 2N2 3: Agricultural Research Station, Winnipeg, MB, Canada R3T 2M9

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