title: Validation Procedure for Multiplex Antibiotic Immunoassays Using Flow-Based Chemiluminescence Microarrays.
abstract: Small molecules like antibiotics or other pharmaceuticals can be detected and quantified, among others, with indirect competitive immunoassays. With regard to multiplex quantification, these tests can be performed as chemiluminescence microarray immunoassays, in which, in principle, the analyte in the sample and the same substance immobilized on the chip surface compete for a limited number of specific antibody binding sites. The amount of the specific primary antibody that has been bound to the surface is visualized by means of a chemiluminescence reaction.Validated quantitative confirmatory methods for the detection of contaminants, for example drug residues, in food samples usually comprise chromatographic analysis and spectrometric detection, e.g., HPLC-MS, GC-MS, or GC with electron capture detection. Here, we describe a validation procedure (according to the Commission Decision of the European Communities 2002/657/EC) for multiplex immunoassays performed as flow-through chemiluminescence microarrays, using the example of a small molecule microarray for the simultaneous detection of 13 antibiotics in milk. By this means, we suggest to accept multianalyte immunoassays as confirmatory methods as well, to benefit from the advantages of a fast automated method that does not need any pretreatment of the sample. The presented microarray chip is regenerable, so an internal calibration is implemented. Therefore, the analytical results are highly precise, combined with low costs (the aim for commercialization is less than 1 € per analyte per sample, this is significantly less than HPLC-MS).
authors.affiliation: Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität of München, Marchioninistraße 17, D-81377, Munich, Germany.
authors.forename: Verena Katharina
authors.initials: VK
authors.lastname: Meyer
authors.affiliation: Instituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, Histopathology and Rapid Tests, Via Bologna 148, I-10154, Turin, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Meloni
authors.affiliation: Instituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, Histopathology and Rapid Tests, Via Bologna 148, I-10154, Turin, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Olivo
authors.affiliation: Chair for Hygiene and Technology of Milk, Veterinary Faculty, Ludwig-Maximilians-Universität München, Schönleutnerstraße 8, D-85764, Oberschleißheim, Germany.
authors.forename: Erwin
authors.initials: E
authors.lastname: Märtlbauer
authors.affiliation: Chair for Hygiene and Technology of Milk, Veterinary Faculty, Ludwig-Maximilians-Universität München, Schönleutnerstraße 8, D-85764, Oberschleißheim, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Dietrich
authors.affiliation: Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität of München, Marchioninistraße 17, D-81377, Munich, Germany.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Niessner
authors.affiliation: Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität of München, Marchioninistraße 17, D-81377, Munich, Germany. michael.seidel@ch.tum.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Seidel
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis*; D002138:Calibration; D007091:Image Processing, Computer-Assisted; D007118:Immunoassay / Q000379:methods*; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000379:methods*; D046228:Microarray Analysis; D012212:Rheology; D012984:Software
chemical_list: D000900:Anti-Bacterial Agents
keywords: Antibiotics; Chemiluminescence; Competitive immunoassay; Flow-based analysis platform; Microarrays; Regeneration; Residue analysis; Screening methods; Veterinary drugs