Competitive Upconversion-Linked Immunosorbent Assay for the Sensitive Detection of Diclofenac

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Authors

HLAVÁČEK Antonín FARKA Zdeněk HUEBNER Maria HORŇÁKOVÁ Veronika NĚMEČEK Daniel NIESSNER Reinhard SKLÁDAL Petr KNOPP Dietmar GORRIS Hans Heiner

Year of publication 2016
Type Article in Periodical
Magazine / Source Analytical Chemistry
MU Faculty or unit

Central European Institute of Technology

Citation
Web http://pubs.acs.org/doi/abs/10.1021/acs.analchem.6b01083
Doi http://dx.doi.org/10.1021/acs.analchem.6b01083
Field Analytic chemistry
Keywords antibody; bioconjugation; diclofenac; immunoassay; pharmaceutical micropollutant; photon-upconversion
Description Photon-upconverting nanoparticles (UCNPs) emit light of shorter wavelength under near-infrared excitation and thus avoid optical background interference. We have exploited this unique photophysical feature to establish a sensitive competitive immunoassay for the detection of the pharmaceutical micropollutant diclofenac (DCF) in water. The so-called upconversion-linked immunosorbent assay (ULISA) was critically dependent on the design of the upconversion luminescent detection label. Silica-coated UCNPs (50 nm in diameter) exposing carboxyl groups on the surface were conjugated to a secondary anti-IgG antibody. We investigated the structure and monodispersity of the nanoconjugates in detail. Using a highly affine anti-DCF primary antibody, the optimized ULISA reached a detection limit of 0.05 ng DCF per mL. This performance came close to a conventional enzyme-linked immunosorbent assay (ELISA) without the need for an enzyme-mediated signal amplification step. The ULISA was further employed for analyzing drinking and surface water samples. The results were consistent with a conventional ELISA as well as liquid chromatography–mass spectrometry (LC–MS).
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