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High-throughput extraction on a dynamic solid phase for low-abundance biomarker isolation from biological samples.

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This study introduces advanced microfluidic fluidized beds for noninvasive liquid biopsies. The new technology efficiently extracts circulating tumor DNA (ctDNA) from serum, improving cancer diagnosis and treatment selection.

Keywords:
EngineeringMicrofluidics

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genomics

Background:

  • Liquid biopsy, particularly circulating tumor DNA (ctDNA) analysis, offers a noninvasive method for cancer management.
  • Efficient extraction of ctDNA from complex biological matrices like serum is critical for accurate analysis.
  • Existing methods face challenges in specificity and minimizing target genetic material loss.

Purpose of the Study:

  • To develop and validate a novel microfluidic fluidized bed system for enhanced ctDNA extraction and preconcentration.
  • To optimize capture efficiency by utilizing flow rate fluctuations and mixed bead sizes.
  • To establish a method for selective capture and detection of mutated BRAF DNA sequences in patient serum.

Main Methods:

  • Development of a new generation of microfluidic fluidized beds (FBs) with a vibration system.
  • Employing a mixture of different bead sizes to enhance bead homogeneity and capture efficiency.
  • Utilizing magnetic FBs for selective capture of double-stranded (dsDNA) BRAF mutated DNA sequences.
  • Ligation chain reaction (LCR) assays for amplification and detection of the target DNA sequence.

Main Results:

  • Demonstrated efficient extraction and preconcentration of specific ctDNA sequences from human serum at flow rates up to 15 µL/min.
  • Showcased significantly enhanced bead homogeneity and capture efficiency through vibration-induced flow rate fluctuations and mixed bead sizes.
  • Successfully developed a method for selective capture of mutated BRAF dsDNA from patient serum.
  • Achieved detection of mutated BRAF DNA sequences at concentrations as low as 6 × 10^4 copies/µL via LCR assays.

Conclusions:

  • The developed microfluidic fluidized bed system offers a high-throughput, efficient, and specific approach for ctDNA extraction from serum.
  • This technology advances noninvasive cancer diagnostics by enabling sensitive detection of specific mutations.
  • The method holds promise for improved cancer diagnosis, treatment selection, and patient monitoring through liquid biopsies.