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Fluorescent identification of immunomagnetically captured CTCs using triplex-aptamer-targeted dendritic SiO2@Fe3O4 nanocomposite

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Abstract

The enumeration of circulating tumor cells (CTCs) in peripheral blood plays a crucial role in the early diagnosis, recurrence monitoring, and prognosis assessment of cancer patients. There is a compelling need to develop an efficient technique for the capture and identification of these rare CTCs. However, the exclusive reliance on a single criterion, such as the epithelial cell adhesion molecule (EpCAM) antibody or aptamer, for the specific recognition of epithelial CTCs is not universally suitable for clinical applications, as it usually falls short in identifying EpCAM-negative CTCs. To address this limitation, we propose a straightforward and cost-effective method involving triplex fluorescently labelled aptamers (FAM-EpCAM, Cy5-PTK7, and Texas Red-CSV) to modify Fe3O4-loaded dendritic SiO2 nanocomposite (dmSiO2@Fe3O4/Apt). This multi-recognition-based strategy not only enhanced the efficiency in capturing heterogeneous CTCs, but also facilitated the rapid and accurate identification of CTCs. The capture efficiency of heterogenous CTCs reached up to 93.33%, with a detection limit as low as 5 cells/mL. Notably, the developed dmSiO2@Fe3O4/Apt nanoprobe enabled the swift identification of captured cells in just 30 min, relying solely on the fluorescently modified aptamers, which reduced the identification time by approximately 90% compared with the conventional immunocytochemistry (ICC) technique. Finally, these nanoprobe characteristics were validated using blood samples from patients with various types of cancers.

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Funding

We appreciated the financial supports from Jiangsu Provincial Key Laboratory of New Drug Research and Clinical Pharmacy Open Research Project (KFKT-2304) and Young Innovative Talent Funding Project (QC202216).

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Authors and Affiliations

  1. Department of Pharmacy, The Affiliated Huaian No.1 People’s Hospital of Nan**g Medical University, 6 Bei**g West Road, Huaian, 223300, Jiangsu, P. R. China

    **nwen Wang, Yu Du, Changchun Cao, **aoli Wu, Kangqun Yang & Liang Zhu

  2. Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, 209 Tongshan Road, Xuzhou, 221004, Jiangsu, P. R. China

    Weijun **g

Authors
  1. **nwen Wang
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  2. Yu Du
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  3. Weijun **g
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  7. Liang Zhu
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Contributions

**nwen Wang: investigation, data curation, writing-original draft. Yu Du: methodology. Weijun **g: visualization, formal analysis. Changchun Cao: TEM, HRTEM, SEM, XRD, VSM, Zeta tests. **aoli Wu: clinical blood sample collections. Kangqun Yang: validation and red blood cell lysis. Liang Zhu: conceptualization, funding acquisition, writing-review and editing.

Corresponding author

Correspondence to Liang Zhu.

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Ethics approval and consent to participate

This study received approval from the Committee on Ethics at the Affiliated Huaian No.1 People’s Hospital of Nan**g Medical University (approval number: YX-Z-2023-034-01), and all participants provided informed consent prior to their participation.

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The authors declare no competing interests.

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Wang, X., Du, Y., **g, W. et al. Fluorescent identification of immunomagnetically captured CTCs using triplex-aptamer-targeted dendritic SiO2@Fe3O4 nanocomposite. Microchim Acta 191, 424 (2024). https://doi.org/10.1007/s00604-024-06504-z

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