Flow cytometric immunoassay for sulfonamides in raw milk

W. de Keizer, M. Bienenmann-Ploum, A.A. Bergwerff, W. Haasnoot

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    Sulfonamide antibiotics are applied in veterinary medicine for the treatment of microbial infections. For the detection of residues of sulfonamides in milk, a multi-sulfonamide flow cytometric immunoassay (FCI) was developed using the Luminex MultiAnalyte Profiling (xMAP) technology. In this automated FCI, a previously developed biotinylated multi-sulfonamide mutant antibody (M.3.4) was applied in combination with fluorescent beads, directly coated with a sulfathiazole derivative, and streptavidin¿phycoerythrin (SAPE) for the detection. With this FCI, at least 11 different sulfonamides could be detected (more than 50% inhibition at the 100 ng mL¿1 level) and, after an incubation of 1 h, measurements were rapid (10 s per sample). For the application with raw milk, a 96-well microplate-based filtration step was included into the protocol to remove disturbing milk fat particles. Because of differences in sensitivity towards different sulfonamides, the FCI was considered and validated as a qualitative screening assay. For sulfadoxine, the most applied sulfonamide in Dutch dairy cattle, the detection capability (CCß) was
    Original languageEnglish
    Pages (from-to)142-149
    Number of pages8
    JournalAnalytica Chimica Acta
    Issue number1-2
    Publication statusPublished - 2008


    • sulfamethazine residues
    • biosensor immunoassay
    • immunobiosensor
    • sulfadiazine
    • serum

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