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Microfluidic platform for negative enrichment of circulating tumor cells

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Abstract

Negative enrichment is the preferred approach for tumor cell isolation as it does not rely on biomarker expression. However, size-based negative enrichment methods suffer from well-known recovery/purity trade-off. Non-size based methods have a number of processing steps that lead to compounded cell loss due to extensive sample processing and handling which result in a low recovery efficiency. We present a method that performs negative enrichment in two steps from 2 ml of whole blood in a total assay processing time of 60 min. This negative enrichment method employs upstream immunomagnetic depletion to deplete CD45-positive WBCs followed by a microfabricated filter membrane to perform chemical-free RBC depletion and target cells isolation. Experiments of spiking two cell lines, MCF-7 and NCI-H1975, in the whole blood show an average of >90 % cell recovery over a range of spiked cell numbers. We also successfully recovered circulating tumor cells from 15 cancer patient samples.

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Abbreviations

CTC:

Circulating tumor cells

WBC:

White blood cell

RBC:

Red blood cell

EMT:

Epithelial to mesenchymal transition

TAC:

Tetrameric antibody complexes

GFP:

Green florescent protein

PBS:

Phosphate buffered saline

EpCAM:

Epithelial cell adhesion molecule

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Acknowledgments

This work was supported by the Science and Engineering Research Council of A*STAR (Agency for Science, Technology and Research), Singapore under the grant number 1031490005. In addition, we greatly appreciate the support from Dr. Chaitanya Kantak for microslit membrane fabrication.

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Authors and Affiliations

  1. BioElectronics Programme, Institute of Microelectronics, A*STAR (Agency for Science, Technology and Research), 1 Science Park Road, Singapore, 117528, Singapore

    Bhuvanendran Nair Gourikutty Sajay, Chia-Pin Chang, Hamizah Ahmad, Chee Chung Wong & Abdur Rub Abdur Rahman

  2. NOVITAS- Nanoelectronics Centre Of Excellence, School of Electrical and Electronics Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798, Singapore

    Bhuvanendran Nair Gourikutty Sajay & Poenar Daniel Puiu

  3. Microfluidics Manufacturing Programme, Singapore Institute of Manufacturing Technology, A*STAR (Agency for Science, Technology and Research), 71 Nanyang Drive, Singapore, 638075, Singapore

    Puttachat Khuntontong & Zhiping Wang

  4. Cancer Science Institute of Singapore, Department of Haematology-Oncology, National University Hospital, 5 Lower Kent Ridge Rd, Singapore, 119074, Singapore

    Ross Soo

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  1. Bhuvanendran Nair Gourikutty Sajay
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  2. Chia-Pin Chang
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  3. Hamizah Ahmad
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  4. Puttachat Khuntontong
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  5. Chee Chung Wong
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  6. Zhiping Wang
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  7. Poenar Daniel Puiu
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  8. Ross Soo
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  9. Abdur Rub Abdur Rahman
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Correspondence to Chia-Pin Chang.

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Bhuvanendran Nair Gourikutty Sajay and Chia-Pin Chang contributed equally to this paper.

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Sajay, B.N.G., Chang, CP., Ahmad, H. et al. Microfluidic platform for negative enrichment of circulating tumor cells. Biomed Microdevices 16, 537–548 (2014). https://doi.org/10.1007/s10544-014-9856-2

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