One-step homogeneous immunoassay for small analytes using recombinant antibodies

Timo Pulli, Matti Höyhtyä, Hans Söderlund, Kristiina Takkinen

    Research output: Contribution to conferenceConference articleScientific

    Abstract

    We have developed a one-step, homogeneous non-competitive immunoassay for small analytes using recombinant antibodies and morphine as the model analyte. Small analytes e.g. abused drugs are an important and increasingly growing area of analytes. Competitive immunoassays performed in microtiter wells or dipsticks are relatively simple methods for detection of drugs of abuse. However, due to the unspecific nature of competitive immunoassays, these tests generate many false positives. Commercial immunoassay tests for morphine and heroin crossreact with e.g. codeine which is a widely used substance in cough medicines. Thus those tests are considered as opiate specific and confirmation of the morphine abuse must always be confirmed by other techniques. This is not only a technical issue but raises also ethical concerns. In our assay format a highly specific antibody against the immune complex (IC) formed between an anti-morphine antibody and morphine was selected from a na¿ve scFv phage display library. The in vitro phage library selection procedure avoids the difficulties associated with the production of anti-IC antibodies by animal immunisation. The anti-morphine and the anti-IC antibodies were labeled with a pair of fluorescence resonance energy transfer (FRET) fluorophores. In the FRET assay the labelled antibodies were incubated with saliva samples spiked with morphine, codeine, or heroin. Within two minutes, 5 ng/ml of morphine, which is clearly under the recommended cutoff level, was detected without cross-reactivity to codeine or heroin. This assay principle is also widely applicable to other small analytes such as pharmaceuticals, toxins, steroids etc.
    Original languageEnglish
    Publication statusPublished - 2006
    MoE publication typeNot Eligible
    Event5th International Congress on Recombinant Antibodies - Zürich, Switzerland
    Duration: 27 Jun 200629 Jun 2006

    Conference

    Conference5th International Congress on Recombinant Antibodies
    Country/TerritorySwitzerland
    CityZürich
    Period27/06/0629/06/06

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