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Fluorescent silver nanoclusters as antibody label in a competitive immunoassay for the complement factor H

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Abstract

Silver nanoclusters (AgNCs) were investigated as labels for the development of a fluoroimmunoassay for the complement factor H (CFH). The reductive one-pot synthesis of AgNCs using lipoic acid as a ligand was optimized by varying the concentration of NaBH4, the temperature and the reaction time. The average diameter and crystal structure of the AgNCs (which display red fluorescence) were determined by HR-TEM. The silver concentration was quantified by ICP-MS. Labelling of the antibody against CFH with AgNCs was optimized. The antibody was labeled with the AgNCs without compromising the recognition capabilities of the antibody. A competitive fluoroimmunoassay was then developed. Fluorescence is measured at excitation/emission maxima of 430/660 nm. The assay has a 0.4 ng mL−1 detection limit and a linear range that extends from 1.2 to 23 ng mL−1. The results compare favorably with those obtained by a commercial ELISA kit. The method was applied to the determination of CFH in spiked human serum.

Schematic presentation of the method for the determination of complement factor H (CFH) protein in human blood by using a CFH-antibody labelled with fluorescent silver nanoclusters.

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Acknowledgements

This work was supported through project CTQ2016-79015-R by Agencia Estatal de Investigación (Spain) and FEDER. B. Fernandez acknowledges her contract RYC-2014-14985 to the Spanish Ministry of Economy and Competitiveness through the “Ramón y Cajal Program”. The Instituto Oftalmológico Fernández-Vega and Fundación de Investigación Oftalmológica acknowledge support from “Cátedra Rafael del Pino” and from Instituto de Desarrollo Económico del Principado de Asturias (IDEPA) and FEDER (project IDE/2016/000214).

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Authors and Affiliations

  1. Department of Physical and Analytical Chemistry, University of Oviedo, Julian Clavería, 8, 33006, Oviedo, Spain

    Eva Valencia, María Cruz-Alonso, Beatriz Fernández & Rosario Pereiro

  2. Instituto Universitario Fernández-Vega (Fundación de Investigación Oftalmológica, Universidad de Oviedo), Avda. Dres. Fernández-Vega, 34, 33012, Oviedo, Spain

    Lydia Álvarez, Héctor González-Iglesias, Beatriz Fernández & Rosario Pereiro

  3. Instituto Oftalmológico Fernández-Vega, Avda. Dres. Fernández-Vega, 34, 33012, Oviedo, Spain

    Héctor González-Iglesias

Authors
  1. Eva Valencia
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  2. María Cruz-Alonso
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  3. Lydia Álvarez
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  4. Héctor González-Iglesias
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Correspondence to Héctor González-Iglesias or Beatriz Fernández.

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Valencia, E., Cruz-Alonso, M., Álvarez, L. et al. Fluorescent silver nanoclusters as antibody label in a competitive immunoassay for the complement factor H. Microchim Acta 186, 429 (2019). https://doi.org/10.1007/s00604-019-3554-y

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