S-Trap, an Ultrafast Sample-Preparation Approach for Shotgun Proteomics

Area of Science:

• Proteomics
• Biochemistry
• Analytical Chemistry

Background:

• Shotgun proteomics success relies heavily on sample preparation.
• Existing methods like filter-aided sample preparation (FASP) are time-consuming and inefficient.

Purpose of the Study:

• To evaluate the performance of the novel Suspension Trapping (S-Trap) filter for shotgun proteomics.
• To compare S-Trap with conventional methods regarding speed, yield, and efficiency.

Main Methods:

• Utilized the S-Trap filter (single and 96-well plate) for protein sample preparation.
• Employed a three-dimensional porous material for protein suspension trapping and in-filter digestion.
• Processed whole-cell lysate and sputum samples.

Main Results:

• S-Trap processing time reduced from ~3 hours (FASP) to <15 minutes.
• Achieved efficient protein trapping, substance depletion, and in-filter digestion.
• Demonstrated comparable global protein identification and quantitation performance.

Conclusions:

• S-Trap represents an ultrafast and effective sample preparation technique for shotgun proteomics.
• The S-Trap filter is a viable alternative to traditional methods, especially for high-throughput applications.
• This method shows promise for clinical sample analysis, such as sputum.