IS-SCP: enhanced single-cell proteomics using an in situ simplified strategy.

Area of Science:

• Proteomics
• Cell Biology
• Biomedical Research

Background:

• Single-cell proteomics allows detailed investigation of individual cell proteomes.
• Existing methods often involve complex sample preparation, leading to sample loss and limited applicability.

Purpose of the Study:

• To develop an advanced, easy-to-use workflow for single-cell proteome analysis.
• To simplify the sample preparation process for enhanced repeatability and depth.

Main Methods:

• Developed an in situ simplified single-cell proteomics (IS-SCP) workflow.
• Optimized reagent mix, volume, and reaction conditions, including a cleavable surfactant (DSSM).
• Simplified pretreatment to a single step of adding samples to a mixed reagent.

Main Results:

• The IS-SCP workflow significantly simplifies single-cell proteome pretreatment.
• Achieved quantification of an average of 3021 protein groups in HeLa cells.
• Achieved quantification of an average of 3289 protein groups in A549 cells.

Conclusions:

• The IS-SCP workflow offers a simplified, repeatable, and deep approach to single-cell proteome analysis.
• This method has potential for investigating cellular heterogeneity at a deep single-cell level.
• Facilitates advanced biomedical research through accessible single-cell proteomic investigations.