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Fluorometric immunoassay for the simultaneous determination of the tumor markers carcinoembryonic antigen and cytokeratin 19 fragment using two kinds of CdSe/ZnS quantum dot nanobeads and magnetic beads

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Abstract

A method is described for the simultaneous determination of the carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1). Two kinds of CdSe/ZnS quantum dot nanobeads (QBs), with emission maxima at 530 nm (green) and 585 nm (yellow), were used as labels, and magnetic beads (MBs) for separation. The MBs were used as substrates to couple CEA and CYFRA21-1 antibody for isolating the proteins. Then, the differently colored QBs were linked to the antibodies against CEA and CYFRA21-1, respectively. Following the formation of the immunocomplex, the intensities of the green and yellow emissions were measured at the same excitation wavelength of 340 nm. The detection limits are 0.1 ng⋅mL− 1 for CEA, and of 0.2 ng⋅mL− 1 for CYFRA21-1. The recoveries from spiked serum are 92.1 - 118.1% for CEA, and from 90.8% to 115.2% for CYFRA21-1, with the relative standard deviations of 6.3 - 12.3% and 7.1 - 11.8%. The method was successfully applied to the simultaneous determination of the two proteins in human serum sample (n = 45). The results correlated well with those of the chemiluminescent enzyme immunoassay kit.

Schematic presentation of the fluorescence immunoassay for the simultaneous determination of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1) based on quantum dot nanobeads (QBs) and magnetic beads (MBs) is reported. The intensities of two kinds of CdSe/ZnS QBs, with the emission maxima at 530 nm (green) and 585 nm (yellow) were measured at the same excitation wavelength of 340 nm.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (81573203, 21605131, 81973099) and Science and Technology Department of Henan Province (172102210006).

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Authors and Affiliations

  1. College of Public Health, Zhengzhou University, Zhengzhou, 450001, China

    Lihua Ding, Xiao Chen, Leiliang He, Fei Yu, Songcheng Yu, Jia Wang, Yongmei Tian, Yilin Wang, Yongjun Wu, Li-e Liu & Lingbo Qu

  2. The Key Laboratory of Nanomedicine and Health Inspection of Zhengzhou, Zhengzhou, 450001, China

    Yongjun Wu & Li-e Liu

  3. College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou, 450001, China

    Lingbo Qu

  4. Henan Joint International Research Laboratory of Green Construction of Functional Molecules and Their Bioanalytical Applications, Zhengzhou University, Zhengzhou, 450001, China

    Lingbo Qu

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  1. Lihua Ding
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Ding, L., Chen, X., He, L. et al. Fluorometric immunoassay for the simultaneous determination of the tumor markers carcinoembryonic antigen and cytokeratin 19 fragment using two kinds of CdSe/ZnS quantum dot nanobeads and magnetic beads. Microchim Acta 187, 171 (2020). https://doi.org/10.1007/s00604-019-3914-7

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