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Related Experiment Videos

High-throughput protein characterization using mass spectrometric immunoassay.

Urban A Kiernan1, Kemmons A Tubbs, Karl Gruber

  • 1Intrinsic Bioprobes, Inc., 625 S. Smith Road Suite, 22, Tempe, Arizona 85281, USA.

Analytical Biochemistry
|January 29, 2002
PubMed
Summary

This study introduces a rapid, high-throughput mass spectrometric immunoassay for analyzing plasma proteins. The system accurately identifies protein modifications and mutations, aiding proteomic and clinical diagnostics.

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Area of Science:

  • Biochemistry
  • Proteomics
  • Analytical Chemistry

Background:

  • Plasma protein analysis is crucial for diagnostics.
  • Existing methods can be time-consuming and complex.
  • High-throughput analysis is needed for large-scale studies.

Purpose of the Study:

  • To develop a rapid, high-throughput system for direct plasma protein analysis.
  • To enable detection of protein modifications and mutations.
  • To facilitate proteomic and clinical diagnostic applications.

Main Methods:

  • Utilized a robotic workstation for antibody-derivatized affinity pipette tips.
  • Employed matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).
  • Screened plasma for transthyretin (TTR) and retinol-binding protein (RBP).

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Main Results:

  • Successfully detected posttranslationally modified TTR and RBP.
  • Identified a mutation in TTR and characterized its nature and site.
  • Achieved rapid analysis at approximately 100 samples per 2 hours.

Conclusions:

  • The developed system offers a rapid and accurate method for plasma protein characterization.
  • This approach is suitable for large-scale proteomic and clinical/diagnostic studies.
  • Enables detailed analysis of protein variants and mutations.