Microfluidic immunocapture of circulating pancreatic cells using parallel EpCAM and MUC1 capture: characterization,...

Area of Science:

• Biomedical Engineering
• Oncology
• Molecular Diagnostics

Background:

• Pancreatic cancer diagnosis often relies on late-stage detection.
• Circulating tumor cells (CTCs) and circulating pancreatic cells (CPCs) are promising biomarkers.
• Current methods for CTC/CPC capture and analysis have limitations.

Purpose of the Study:

• To develop and optimize a microfluidic device for efficient capture and analysis of CPCs and CTCs.
• To enable downstream genetic analysis of captured cells for mutation detection.
• To establish a platform for early pancreatic cancer diagnosis and risk stratification.

Main Methods:

• Microfluidic device with parallel anti-EpCAM and MUC1 immunocapture.
• Detection of KRAS mutation using RT-PCR and Sanger sequencing.
• Targeted membrane lysis for intact nuclei isolation and single-cell genetic analysis.
• Staining protocol with CTC markers (DAPI, CK, CD45) and MUC4.
• Semi-automated image analysis for CPC identification.

Main Results:

• Efficient capture of CPCs and CTCs using combined immunocapture.
• Successful detection of an oncogenic KRAS mutation in spiked samples.
• Demonstrated feasibility of single-cell genetic analysis.
• Developed a comprehensive staining and analysis protocol.
• Initial clinical sample analysis suggests improved capture of population heterogeneity.

Conclusions:

• The developed microfluidic platform enables efficient capture and molecular analysis of CPCs and CTCs.
• This technology holds potential for early pancreatic cancer detection and patient risk stratification.
• Further validation in clinical settings is warranted to establish its diagnostic utility.