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Homogeneous immunoassay for the cancer marker alpha-fetoprotein using single wavelength excitation fluorescence cross-correlation spectroscopy and CdSe/ZnS quantum dots and fluorescent dyes as labels

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Abstract

The article describes sensitive and selective homogeneous immunoassays for the liver cancer biomarker alpha-fetoprotein (AFP) in human serum by using single wavelength excitation fluorescence cross-correlation spectroscopy (SW-FCCS). Both competitive and sandwich immunoassay modes were applied, and AFP served as a model analyte. Fluorescent CdSe/ZnS quantum dots (with a 655 nm emission peak) and the fluorophore Alexa Fluor 488 (520 nm emission) were chosen to label the antibodies in the sandwich mode, and the antibody and the antigen in the competitive mode. Under optimized conditions, the sandwich assay has a linear dynamic range that covers the 20 pM to 5.0 nM concentration range. The competitive assay, in turn, extends from 180 pM to 15.0 nM. The respective detection limits are 20 pM and 180 pM. The method was successfully applied to directly determine AFP in (spiked) clinical samples, and results were in good agreement with data obtained via ELISAs.

Homogeneous immunoassay based on single wavelength excitation fluorescence cross-correlation spectroscopy (SW-FCCS) has been used to detect the biomarker alpha-fetoprotein in human sera.

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Acknowledgments

This work was financially supported by NSFC (Grants 21075081, 20905048, 21135004 and 21327004).

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Authors and Affiliations

  1. College of Chemistry & Chemical Engineering, State Key Laboratory of Metal Matrix Composites, Shanghai Jiaotong University, 800 Dongchuan Road, Shanghai, 200240, People’s Republic of China

    Jinjie Wang, Heng Liu, Xiangyi Huang & Jicun Ren

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  1. Jinjie Wang
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  2. Heng Liu
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  3. Xiangyi Huang
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  4. Jicun Ren
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Correspondence to Xiangyi Huang or Jicun Ren.

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Wang, J., Liu, H., Huang, X. et al. Homogeneous immunoassay for the cancer marker alpha-fetoprotein using single wavelength excitation fluorescence cross-correlation spectroscopy and CdSe/ZnS quantum dots and fluorescent dyes as labels. Microchim Acta 183, 749–755 (2016). https://doi.org/10.1007/s00604-015-1694-2

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  • DOI: https://doi.org/10.1007/s00604-015-1694-2

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