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Improving Proteome Coverage and Sample Recovery with Enhanced FASP (eFASP) for Quantitative Proteomic Experiments
Jonathan Erde1, Rachel R Ogorzalek Loo2, Joseph A Loo3,4
1Department of Chemistry and Biochemistry, University of California-Los Angeles, Los Angeles, CA, 90095, USA.
Methods in Molecular Biology (Clifton, N.J.)
|February 12, 2017
Summary
Enhanced Filter Aided Sample Preparation (eFASP) improves proteomics by reducing sample loss and increasing hydrophobic protein detection. This method yields clean protein digests ready for direct LC-MS analysis.
Area of Science:
- Proteomics
- Biochemistry
- Analytical Chemistry
Background:
- Traditional Filter Aided Sample Preparation (FASP) can lead to sample loss.
- Hydrophobic proteins are often underrepresented in proteomic analyses.
- Contaminants can interfere with downstream mass spectrometry analysis.
Purpose of the Study:
- To develop an improved sample preparation method for proteomics.
- To minimize sample loss during protein digestion.
- To enhance the detection of hydrophobic proteins.
Main Methods:
- The enhanced Filter Aided Sample Preparation (eFASP) method was developed.
- Plastics passivation techniques were incorporated.
- Digestion-enhancing surfactants were utilized within the FASP workflow.
Main Results:
- The eFASP method significantly reduced sample loss compared to traditional FASP.
- Increased representation of hydrophobic proteins was observed in eFASP samples.
- Protein digests generated by eFASP were free of common contaminants.
Conclusions:
- eFASP is an effective method for improving protein sample preparation in proteomics.
- The method enhances both qualitative and quantitative proteomic analyses.
- eFASP facilitates direct analysis of protein digests by LC-MS, streamlining workflows.
