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Proteomic Sample Preparation from Formalin Fixed and Paraffin Embedded Tissue
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Pressure-assisted tryptic digestion using a syringe.

Hyo-Jik Yang1, Jangmi Hong, Sunyoung Lee

  • 1Department of Chemistry, Chungnam National University, Daejeon, South Korea.

Rapid Communications in Mass Spectrometry : RCM
|March 3, 2010
PubMed
Summary
This summary is machine-generated.

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Researchers developed a rapid protein digestion method using a syringe to apply pressure. This pressure-assisted tryptic digestion significantly improves efficiency, yielding more peptides in just 30 minutes compared to overnight methods.

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Efficient protein digestion is crucial for mass spectrometry-based proteomics.
  • Traditional methods can be time-consuming, often requiring overnight incubation.
  • Optimizing digestion protocols can enhance peptide identification and sequence coverage.

Purpose of the Study:

  • To develop a simple, rapid, and effective method for tryptic protein digestion.
  • To investigate the impact of pressure application on digestion efficiency.
  • To compare pressure-assisted digestion with conventional atmospheric pressure methods.

Main Methods:

  • Utilized a 3 mL syringe to apply 6 atm pressure during tryptic digestion.
  • Tested the method on three model proteins: cytochrome c, horse heart myoglobin, and bovine serum albumin (BSA).

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Evaluation of Protein&#8211;Protein Interactions using an On-Membrane Digestion Technique
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Related Experiment Videos

Last Updated: Jun 15, 2026

Proteomic Sample Preparation from Formalin Fixed and Paraffin Embedded Tissue
09:20

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Published on: September 2, 2013

A Spin-Tip Enrichment Strategy for Simultaneous Analysis of N-Glycopeptides and Phosphopeptides from Human Pancreatic Tissues
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A Spin-Tip Enrichment Strategy for Simultaneous Analysis of N-Glycopeptides and Phosphopeptides from Human Pancreatic Tissues

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Evaluation of Protein&#8211;Protein Interactions using an On-Membrane Digestion Technique
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Evaluation of Protein–Protein Interactions using an On-Membrane Digestion Technique

Published on: July 19, 2019

  • Analyzed digested peptides using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS).

    • Main Results:

    • Complete disappearance of protein peaks in MALDI-MS spectra within 30 min at 6 atm and 37°C.
    • Pressure-assisted digestion yielded a greater number of peptides compared to overnight atmospheric digestion.
    • Achieved comparable or improved sequence coverage for model proteins in 30 min versus overnight digestion.

    Conclusions:

    • Syringe-based pressure application is an effective strategy to accelerate tryptic digestion.
    • This method offers a significant time-saving advantage without compromising peptide yield or sequence coverage.
    • The developed technique provides a simple and efficient alternative for protein sample preparation in proteomics.