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

Hannah M Pezzi1, David J Guckenberger, Jennifer L Schehr

  • 1Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, 1111 Highland Avenue, Madison, Wisconsin 53705, USA.

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|October 19, 2018
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Summary
This summary is machine-generated.

The multiplexed technology for automated extraction (mTAE) platform efficiently isolates rare cells, like circulating tumor cells (CTCs), from patient samples. This versatile technology enables downstream molecular analysis for disease monitoring.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Oncology

Background:

  • Rare cell populations are crucial for monitoring disease progression and treatment response.
  • Isolating rare cells is challenging due to non-target cell variability and disease-related changes.
  • Cell isolation platforms need to be versatile, efficient, and pure across diverse sample types.

Purpose of the Study:

  • To evaluate the multiplexed technology for automated extraction (mTAE) as a versatile platform for rare cell isolation.
  • To demonstrate the capability of mTAE in isolating circulating tumor cells (CTCs) from cancer patient samples.
  • To showcase the integrated functionalities of mTAE, including protein staining and nucleic acid extraction.

Main Methods:

  • Utilized a magnetic bead-based isolation platform (mTAE) supporting positive, negative, and combinatorial selection strategies.
  • Validated mTAE by isolating circulating tumor cells (CTCs) from breast and prostate cancer patient blood samples.
  • Performed on-platform protein staining and nucleic acid isolation following cell capture.

Main Results:

  • Negative selection demonstrated high efficiency for CTC capture.
  • Positive selection achieved higher purity, with an average of 95 contaminant cells/mL.
  • Combinatorial selection showed an increased overall capture efficiency.
  • Detected androgen receptor-related transcripts from CTCs isolated from prostate cancer patients.

Conclusions:

  • The mTAE platform offers flexibility in selection strategies (negative, positive, combinatorial) for rare cell isolation.
  • Integrated capabilities of mTAE (isolation, staining, nucleic acid extraction) facilitate comprehensive interrogation of rare cell populations.
  • mTAE's adaptability is essential for studying heterogeneous rare cell populations in clinical samples.