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Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
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Integrated device for online sample buffer exchange, protein enrichment, and digestion.

Liangliang Sun1, Junfeng Ma, Xiaoqiang Qiao

  • 1National Chromatographic R. & A. Center, Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Science, Dalian 116023, China.

Analytical Chemistry
|February 16, 2010
PubMed
Summary
This summary is machine-generated.

A novel integrated device simplifies proteomic analysis by combining buffer exchange, protein enrichment, and enzymatic digestion. This automated system significantly reduces sample preparation time, enabling high-throughput proteome analysis.

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Last Updated: Jun 16, 2026

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Proteomics

Background:

  • Traditional proteomic sample preparation is time-consuming and labor-intensive.
  • Online integration of sample preparation steps can improve efficiency and reduce errors.

Purpose of the Study:

  • To develop an integrated sample treatment device for simultaneous buffer exchange, protein enrichment, and online enzymatic digestion.
  • To evaluate the device's performance for proteomic analysis coupled with liquid chromatography and mass spectrometry.

Main Methods:

  • Development of a device integrating a membrane interface and a hybrid silica-based immobilized enzymatic reactor (IMER).
  • Online processing of protein eluates, including buffer exchange and trypsin digestion.
  • Analysis of protein digests using micro-liquid chromatography-electrospray ionization-tandem mass spectrometry (microLC-ESI-MS/MS).

Main Results:

  • The integrated device effectively reduced acetonitrile content and adjusted pH for optimal trypsin digestion.
  • Signal intensity of myoglobin digests was enhanced over 10-fold.
  • The device achieved comparable results to traditional offline methods but reduced processing time by 192-fold.

Conclusions:

  • The developed integrated sample treatment device offers a highly efficient solution for proteomic sample preparation.
  • This technology has the potential for online coupling with separation and identification techniques for high-throughput proteome analysis.