A monoclonal antibody to aflatoxin B1: detection of the mycotoxin by enzyme immunoassay |
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| Authors: | A AG Candlish W H Stimson J E Smith |
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| Institution: | Division of Immunology, Department of Bio science and Biotechnology, University of Strathclyde, Taylor Street, Glasgow, UK;*Division of Applied Microbiology, Department of Bioscience and Biotechnology, University of Strathclyde, George Street, Glasgow, UK |
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| Abstract: | A monoclonal antibody (McAb) was produced after fusion of mouse (X63.Ag8.6.5.3) myeloma cells with spleen cells isolated from female Balb-c/NZB F1 hybrid mice immunized with aflatoxin B1 (oxine)-keyhole limpet haemocyanin conjugate. The hybridoma cell line producing antibody specific for aflatoxin B1 (AFB1) was grown in tissue culture and as an ascites tumour. The ascitic fluid gave suitably high dilution titres (1:800 000) by enzyme immunoassay and was conjugated to horseradish peroxidase by a two-step procedure with glutaraldehyde. The conjugate was used to develop a direct competitive enzyme-linked immunosorbent (ELISA) assay for AFB1. The sensitivity of the ELISA was 0–2 ng/ml with a working range up to 10 ng/ml for AFB1. The specificity of the McAb was determined and it was shown not to cross-react significantly with any of the metabolites tested. This McAb and the direct competitive ELISA described may prove of use in the detection of AFB1 in foods and feeds. |
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