A Multifunctional Platform for the Capture, Release, And Enumeration of Circulating Tumor Cells Based on Aptamer Binding, Nicking Endonuclease-Assisted Amplification, And Inductively Coupled Plasma Mass Spectrometry Detection.

Abstract

Inductively coupled plasma mass spectrometry (ICP-MS) combined with element tags has been well designed and extensively applied in cell enumeration. It possesses superior quantitative capability, strong resistance to matrix interference, multiplex detection capability but destructive characteristic. Herein, we constructed an ICP-MS based multifunctional platform for capture, nondestructive enumeration, and release of circulating tumor cells (CTCs). Aptamer on capture probe recognized Mucin 1 (MUC1) on membrane of MCF-7 cells specifically, thus the cells were captured by probe and the Initiator originally hybridized with Aptamer was substituted by MUC1 and released into solution. Then the released Initiator was separated from the captured cells and hybridized with Tb labeled Substrate on detection probe to release a large amount of nicked Tb fragments through the nicking endonuclease assisted amplification for subsequent ICP-MS detection. Meanwhile, cells captured by probe were released by nuclease digestion for further reculture. Such a strategy effectively avoids CTCs destruction resulted from ICP-MS enumeration, increases the detection sensitivity of ICP-MS by involving nicking endonuclease assisted amplification, and realizes cell recovery for further analysis. A limit of detection of 87 MCF-7 cells and a linear range of 250-10 000 MCF-7 cells were realized for ICP-MS enumeration. A cell recovery of 52.7% (with capture and release efficiency of 63.9 and 82.5%, respectively) and a viability of 74.3% were obtained, meanwhile the released cells exhibited strong proliferation ability. This multifunctional platform for CTCs capture, enumeration, and release has great applicable potential in early diagnosis and individual treatment for cancer.

• Authors: Xiao Yin, Beibei Chen, Man He, Bin Hu
• Journal: Analytical chemistry (Anal Chem) Vol. 92 Issue 15 Pg. 10308-10315 (08 04 2020) ISSN: 1520-6882 [Electronic] United States
• PMID: 32615753 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Aptamers, Nucleotide
• Endonucleases

Topics

• Aptamers, Nucleotide (chemistry)
• Cell Count (methods)
• Cell Survival
• Endonucleases (chemistry, metabolism)
• Humans
• MCF-7 Cells
• Mass Spectrometry (methods)
• Neoplastic Cells, Circulating (chemistry)
• Protein Binding