4.8 Article

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

Journal

ANALYTICAL CHEMISTRY
Volume 88, Issue 11, Pages 6011-6017

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.6b01083

Keywords

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Funding

  1. COST Action The European Upconversion Network: From the Design of Photon-Upconverting Nanomaterials to Biomedical Applications [CM1403]
  2. German Research Foundation (DFG) [1968/5-1]
  3. Program of Employment of Newly Graduated Doctors of Science for Scientific Excellence [CZ.1.07/2.3.00/30.0009]
  4. Czech Ministry of Education, Youth and Sports (COST CZ Project) [LD15023]
  5. ANR-DFG program (Project NArBioS) [ANR-11-INTB-1013]
  6. German Academic Exchange Service (DAAD)
  7. Grant Agency of the Czech Republic [14-28254S]
  8. Agence Nationale de la Recherche (ANR) [ANR-11-INTB-1013] Funding Source: Agence Nationale de la Recherche (ANR)

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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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