Label-free single cell proteomics utilizing ultrafast LC and MS instrumentation: A valuable complementary technique...

Area of Science:

• Proteomics
• Genomics
• Single-cell analysis

Background:

• Understanding gene expression's translation to phenotype requires mapping proteomic data to transcriptomic data.
• Unlike nucleic acid sequencing, in vitro protein amplification is impossible, and no gold standard single-cell proteomic workflow exists.
• Advances in microfluidics, separation, data acquisition, and analysis have improved tiny sample analysis.

Purpose of the Study:

• To review recent advances in label-free single-cell mass spectrometry (SCP) workflows.
• To guide researchers in selecting optimal SCP methods for their applications.
• To highlight future potential and limitations of SCP techniques.

Main Methods:

• Focus on label-free single-cell mass spectrometry (SCP) workflows.
• Discussion of microfluidic sample preparation, multi-dimensional separation, data acquisition, and analysis.
• Evaluation of multiplexing versus label-free strategies for sensitivity, robustness, and throughput.

Main Results:

• Label-free SCP is a promising strategy for high dynamic range and unbiased quantification.
• Recent advancements have significantly improved performance in analyzing small sample amounts.
• Sensitivity, robustness, and throughput remain critical needs in the field.

Conclusions:

• Label-free SCP workflows are advancing rapidly, offering new possibilities for single-cell proteomics.
• Careful method selection is crucial for specific applications.
• Future research should address current limitations and explore new applications.