Sickle-like Inertial Microfluidic System for Online Rare Cell Separation and Tandem Label-Free Quantitative...

Area of Science:

• Biotechnology
• Proteomics
• Microfluidics

Background:

• Label-free proteomics offers valuable insights for personalized medicine.
• Low abundance of rare clinical cells, like circulating tumor cells (CTCs), hinders conventional proteomic analysis.
• Need for advanced techniques to analyze rare cells in trace clinical samples.

Purpose of the Study:

• To develop a system for simultaneous separation and label-free proteomic analysis of rare cells.
• To enable personalized medicine through insights from trace clinical samples.
• To overcome the limitations of analyzing low-abundance clinical samples.

Main Methods:

• Development of a sickle-like inertial microfluidic system for online rare cell separation.
• Integration of tandem label-free proteomics (Orcs-proteomics) with a specific buffer system for cell lysis and reductive alkylation.
• Demonstration using 293T cells, simulated patient blood samples (MCF7 cells spiked with white blood cells), and actual patient CTCs.

Main Results:

• Orcs-proteomics successfully identified thousands of protein groups from varying numbers of 293T cells (913 to 2770 proteins from 4 to 119 cells).
• Over 2000 protein groups were identified from an average of 61 MCF7 cells spiked with white blood cells.
• Proteomic analysis of 5-7 circulating tumor cells (CTCs) from advanced breast cancer patients yielded 973 and 1135 protein groups, respectively.

Conclusions:

• Orcs-proteomics effectively separates and quantifies proteins from rare cells in trace clinical samples.
• This technology provides crucial technical support for personalized treatment decision-making using rare primary patient samples.
• Enables deeper understanding of disease mechanisms and biomarker discovery from minimal biological material.