Versatile exclusion-based sample preparation platform for integrated rare cell isolation and analyte extraction

Hannah M. Pezzi, David J. Guckenberger, Jennifer L. Schehr, Jacob Rothbauer, Charlotte Stahlfeld, Anupama Singh, Sacha Horn, Zachery D. Schultz, Rory M. Bade, Jamie M. Sperger, Scott M. Berry, Joshua M. Lang, David J. Beebe

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Rare cell populations provide a patient-centric tool to monitor disease treatment, response, and resistance. However, understanding rare cells is a complex problem, which requires cell isolation/purification and downstream molecular interrogation-processes challenged by non-target populations, which vary patient-to-patient and change with disease. As such, cell isolation platforms must be amenable to a range of sample types while maintaining high efficiency and purity. The multiplexed technology for automated extraction (mTAE) is a versatile magnetic bead-based isolation platform that facilitates positive, negative, and combinatorial selection with integrated protein staining and nucleic acid isolation. mTAE is validated by isolating circulating tumor cells (CTCs)-a model rare cell population-from breast and prostate cancer patient samples. Negative selection yielded high efficiency capture of CTCs while positive selection yielded higher purity with an average of only 95 contaminant cells captured per milliliter of processed whole blood. With combinatorial selection, an overall increase in capture efficiency was observed, highlighting the potential significance of integrating multiple capture approaches on a single platform. Following capture (and staining), on platform nucleic acid extraction enabled the detection of androgen receptor-related transcripts from CTCs isolated from prostate cancer patients. The flexibility (e.g. negative, positive, combinatorial selection) and capabilities (e.g. isolation, protein staining, and nucleic acid extraction) of mTAE enable users to freely interrogate specific cell populations, a capability required to understand the potential of emerging rare cell populations and readily adapt to the heterogeneity presented across clinical samples.

Original languageEnglish
Pages (from-to)3446-3458
Number of pages13
JournalLab on a Chip
Volume18
Issue number22
DOIs
StatePublished - Nov 21 2018

Bibliographical note

Funding Information:
We would like to thank all patients who participated in this study. We are also grateful for the help of the UWCCC clinical research group, especially Jamie Wiepz, Kelly Bush, Amy For-syth, Dorothea Horvath, Jane Straus, Mary Jane Staab, Dr. Glenn Liu, Dr. Douglas McNeel, Dr. Christos Kyriakopolous, Dr. C. Pettaway, and Dr. George Wilding. This work was supported by a Movember-Prostate Cancer Foundation Challenge Award (to J. M. Lang and D. J. Beebe), the Bill & Melinda Gates Foundation through the Grand Challenges in Global Health Initiative (to S. M. Berry and D. J. Beebe), NIH grant #1R01CA181648 (to S. M. Berry and J. M. Lang), Department of Defense PCRP grant #W81XWH-12-1-0052 (to J. M. Lang), Department of Defense Synergistic Idea Development Award BC 150425 (to J. M. Lang), NIH grant #5R33CA137673 (to D. J. Beebe), University of Wisconsin Carbone Cancer Center Support Grant NIH P30 (to D. J. Beebe, S. M. Berry, D. J. Guckenberger) and NSF GRFP DGE-0718123 (to D. J. Beebe).

Publisher Copyright:
© The Royal Society of Chemistry.

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Chemistry (all)
  • Biomedical Engineering

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