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Multiplex approach for an immunological detection of drugabuse

A validation study
: Schumacher, Sarah; Seitz, Harald

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Toxichem Krimtech 39 (2015), No.82, Special Issue, pp.151-154
ISSN: 2190-3441
Gesellschaft für Toxikologische und Forensische Chemie (GTFCh Symposium) <29, 2015, Mosbach>
Journal Article, Conference Paper, Electronic Publication
Fraunhofer IZI ()

Aim: The aim of this study was to develop a multiplex immunoassay for nine different drugs. Therefore each reagent undergoes a stringent quality control e.g. antibodies used has to be validated with at least 2 independent methods.
Methods: For validation Western Blot analysis and ELISA were performed. A competitive ELISA was established allowing the quantification of the drugs in sera. Appropriate controls were included for background subtraction, determination of unspecific signals and assay control. Furthermore the assay was miniaturized on a microarray, which was produced with a non-contact spotter.
Results: For 4 out of 9 selected drugs specific antibodies could be obtained and a competitive ELISA established for quantification. Validated antibodies were characterized by no cross-reactivity to serum and no unspecific binding to other compounds. Serum samples with spiked drugs or samples from the LKA Berlin were analyzed. Each sample was performed in triplicates and each experiment was done twice at least. Limits of quantification meet the requirements of the GTFCh. First multiplex and spotting experiments were done with a satisfying result.
Conclusion: The presented approach enables a sensitive and reliable detection method for drug abuse in sera. The validation studies are continued for the remaining drugs.