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Mycotoxins particularly afl atoxins are gaining increasing importance due to their deleterious effects on human and animal health and also due to ubiquitous presence of afl atoxigenic fungi in all the agricultural products. Corn, groundnuts and other plants were infected by Aspergillus fl avus & Aspergillus parasiticus and secrete the mycotoxins. Afl atoxin B1 transmitted to cow by feeding these infected plants and afl atoxin B1 transformed into its hydroxylated product such as afl atoxin M1 and M2 and such afl atoxin secreted in cow milk. In the present study, we explored the nanobiotechnology approach to prepare detection bioconjugates of gold coated iron oxide nanoparticle for the detection of afl atoxin M1 in milk. Gold coated iron oxide nanoparticles (Au-Fe3O4 NPs) ranging between 10-20 nm were synthesized by co-precipitation method and functionalized by EDC-NHS then labeled with FITC-labeled streptavidin (FITC-STV). Biotinylated afl atoxin M1 (biAFM1) specifi c to monoclonal anti-afl atoxin M1 antibodies (mAFM1) were prepared, separately. The fl uorophore FITC-STV-Au-Fe3O4 NPs and bimAFM1 antibodies were allowed to interact to obtain Au-Fe3O4 NPs-FITC-STV-bimAFM1 antibody and AFM1 bioconjugates. The bioconjugates were characterized by Transmission Electron Microscope (TEM), Scanning Electron Microscope with Energy Dispersive X-ray (SEM-EDAX), Fourier Transform Infrared (FTIR), Particle Size Analyzer with zeta potential (PSA) and Fluorescence Microscopy. The bioconjugation formation was confi rmed by pinkish red color in bright fl ied microscopy same fi eld observed by fl uorescence microscopy shows the green fl uorescence which confi rm the aggregation between Au-Fe3O4 NPs-FITC labeled STV-bimAFM1 antibody and AFM1.

Formulation of detection bioconjugates of gold coated iron oxide nanoparticles for afl atoxin M1 in milk