The purpose of the article is to establish a quick enrichment and detection method using immunomagnetic beads and flow cytometry to analyze circulating tumor cells (CTCs) in the peripheral blood.
After incubation with CD326-PE and CD45-APC antibodies, more than 60% MCF7 cells in M-Buffer could be detected while less than 10% of the same cells could be detected by flow cytometry (FCM) if spiked into blood. However, in combination with CD326 and CD45 immunomagnetic beads, detection rate of MCF7 cells in blood reached 57%. For circulating tumor cells, enrichment by CD326 and CD45 immunomagnetic beads improve the detection rate from nearly undetectable to more than 24.14%.
Live CTCs in peripheral blood can be effectively and sensitively detected by using a combination of immunomagnetic beads (CD45 and CD326) and flow cytometry.
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This work was supported by the 100-Talent Program of Chinese Academy of Sciences, the National Natural Science Foundation of China (Nos. 81572948, 21772201, 21703254), the co-operative grant from Anhui Medical University and Center of Medical Physics and Technology (No. LHJJ202007).
Conflict of interest
All authors declare that they have no conflict of interest.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
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Lei Hu and Xueran Chen have contributed equally to this work.
Electronic supplementary material
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Supplementary Figure 1—Enriched CTC could be effectively detected by flow cytometry. Four venous blood species from Patient 4-7 with malignant tumors were collected. 20 µL of each 5 mL collected blood sample were immediately incubated with CD45-APC and CD326-PE antibodies while the rest samples sequential underwent CD45 and CD326 immunomagnetic beads enrichment followed by fluorescence antibodies label. All four groups (un-enriched, CD45+, CD326- and CD326+) were harvested by 300g centrifuge at 4 °C, 15min and detected by FCM. Supplementary file 1 (TIF 3759 kb)
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Hu, L., Chen, X., Chen, M. et al. Enrichment and detection of circulating tumor cells by immunomagnetic beads and flow cytometry. Biotechnol Lett (2020). https://doi.org/10.1007/s10529-020-03007-8
- Circulating tumor cells
- Flow cytometry
- Fluorescence antibodies
- Immunomagnetic beads