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A Magnetic Nanoparticle-Based Time-Resolved Fluoroimmunoassay for Determination of the Cytokeratin 19 Fragment in Human Serum

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Abstract

A sensitive, rapid and novel measurement method for cytokeratin 19 fragment (CYFRA 21–1) in human serum by magnetic particle-based time-resolved fluoroimmunoassay (TRFIA) is described. Built on a sandwich-type immunoassay format, analytes in samples were captured by one monoclonal antibody coating onto the surface of magnetic beads and “sandwiched” by another monoclonal antibody labeled with europium chelates. The coefficient variations of the method were lower than 7 %, and the recoveries were in the range of 90–110 % for serum samples. The lower limit of quantitation of the present method for CYFRA 21–1 was 0.78 ng/ml. The correlation coefficient of CYFRA 21–1 values obtained by our novel TRFIA and CLIA was 0.980. The present novel TRFIA demonstrated high sensitivity, wider effective detection range and excellent reproducibility for determination of CYFRA 21–1 can be useful for early screening and prognosis evaluation of patients with non-small cell lung cancer.

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Abbreviations

NSCLC:

Non-small cell lung cancer

CYFRA 21–1:

Cytokeratin 19 fragment

TRFIA:

Time-resolved fluoroimmunoassay

BSA:

Ovine serum albumin

MES:

4-morpholineethanesulfonic acid

NHS:

N-hydroxysulfosuccinimide

EDC:

1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride

Eu:

Europium

CLIA:

Chemiluminescence immunoassay

McAb:

Monoclonal antibody

LLOQ:

Lower limit of quantitation

RE:

Relative error

CV:

Coefficient of variation

SD:

Standard deviation

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Acknowledgments

The work was supported by the National Natural Science Foundation of China (Grant No. 81271931) and the Natural Science Foundation of Guangdong Province (Grant No. S2012010009547).

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Authors and Affiliations

  1. Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou, China

    Guanfeng Lin, Tiancai Liu, Jingyuan Hou, Zhiqi Ren, Jianwei Zhou, Qianni Liang, Zhenhua Chen & Yingsong Wu

  2. Department of Biopharmaceutical, School of Biotechnology, Southern Medical University, Guangzhou, China

    Wenqi Dong

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  1. Guanfeng Lin
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Correspondence to Wenqi Dong or Yingsong Wu.

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Lin, G., Liu, T., Hou, J. et al. A Magnetic Nanoparticle-Based Time-Resolved Fluoroimmunoassay for Determination of the Cytokeratin 19 Fragment in Human Serum. J Fluoresc 25, 361–367 (2015). https://doi.org/10.1007/s10895-015-1518-0

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  • DOI: https://doi.org/10.1007/s10895-015-1518-0

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