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Related Experiment Video

Updated: Jul 14, 2026

Fast Enzymatic Processing of Proteins for MS Detection with a Flow-through Microreactor
09:49

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Published on: April 6, 2016

Gold nanoparticle assembly microfluidic reactor for efficient on-line proteolysis.

Yun Liu1, Yan Xue, Ji Ji

  • 1Department of Chemistry and Institute of Biomedical Sciences, Fudan University, Shanghai 200433, China.

Molecular & Cellular Proteomics : MCP
|May 24, 2007
PubMed
Summary

Researchers developed a novel microchip reactor using gold nanoparticles to efficiently digest low-level proteins from mouse macrophages. This gold nanoparticle network entrapping trypsin enables sensitive protein analysis and identification.

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Area of Science:

  • Biochemistry
  • Nanotechnology
  • Analytical Chemistry

Background:

  • Efficient on-line proteolysis is crucial for analyzing low-level proteins in complex biological samples.
  • Existing methods often face challenges with sensitivity and sample complexity, particularly for macrophage extracts.

Purpose of the Study:

  • To design and characterize a microchip reactor for efficient on-line proteolysis of trace proteins.
  • To develop a nanostructured surface coating using gold nanoparticles for enzyme immobilization.
  • To assess the bioactivity and performance of entrapped trypsin for protein digestion.

Main Methods:

  • Assembly of a nanostructured surface coating via layer-by-layer electrostatic binding of poly(diallyldimethylammonium chloride) and gold nanoparticles.
  • Monitoring of the assembly process using UV-visible spectroscopy, atomic force microscopy, and quartz crystal microbalance.

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Last Updated: Jul 14, 2026

Fast Enzymatic Processing of Proteins for MS Detection with a Flow-through Microreactor
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Published on: April 6, 2016

Millifluidics for Chemical Synthesis and Time-resolved Mechanistic Studies
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Aqueous Droplets Used as Enzymatic Microreactors and Their Electromagnetic Actuation
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  • Theoretical study of trypsin adsorption using the Langmuir isotherm model and enzymatic kinetics assays.

    • Main Results:

    • A microchip reactor with a gold nanoparticle network entrapping trypsin was successfully fabricated.
    • The entrapped trypsin demonstrated preserved bioactivity with a maximum proteolytic rate of 400 mM/(min.microg).
    • The reactor efficiently digested trace amounts of proteins down to femtomole levels, enabling identification by MALDI-TOF MS/MS and LC-ESI-MS/MS.

    Conclusions:

    • The developed gold nanoparticle-coated microchip reactor provides an efficient platform for on-line proteolysis of low-level proteins.
    • This method allows for sensitive and comprehensive protein identification from complex biological samples like mouse macrophage extracts.
    • The nanostructured coating effectively immobilizes trypsin while maintaining its enzymatic activity.