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Related Experiment Video

Updated: Jul 8, 2026

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
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Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

Published on: August 17, 2015

ABRF ESRG 2006 study: Edman sequencing as a method for polypeptide quantitation.

D C Brune1, B Hampton, R Kobayashi

  • 1Dept. of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287-1604, USA. dbrune@asu.edu

Journal of Biomolecular Techniques : JBT
|January 2, 2008
PubMed
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This summary is machine-generated.

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This study evaluated Edman sequencing accuracy for protein analysis, finding high accuracy in amino acid identification and favorable quantitative results compared to mass spectrometry.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Proteomics

Background:

  • Edman degradation is a key method for protein and peptide sequencing.
  • Inter-laboratory comparisons are vital for validating analytical services.

Purpose of the Study:

  • To assess the accuracy of Edman sequencing for quantitative polypeptide analysis.
  • To identify a modified amino acid, N-epsilon-acetyl lysine [Lys(Ac)], in synthetic peptides.

Main Methods:

  • A mixture of three synthetic peptides (B, C, and C* with Lys(Ac)) was analyzed by participating laboratories.
  • Participants performed Edman sequencing, collecting data on amino acid assignments, yields, and peak areas.
  • Mass spectrometry data (MALDI-TOF, ESI) was collected for comparative analysis.

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Last Updated: Jul 8, 2026

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
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Main Results:

  • Edman sequencing achieved 88-93% accuracy in amino acid assignments for peptides B and C.
  • Average initial sequencing yields were 67% for peptide (C+C*) and 65.6% for peptide B.
  • Relative molar ratios determined by Edman sequencing compared favorably with mass spectrometry, despite a 49% error in Lys(Ac) quantification due to standard unavailability.

Conclusions:

  • Edman sequencing demonstrates high accuracy for both qualitative and quantitative peptide analysis.
  • The study highlights the reliability of Edman sequencing for protein analysis, even with modified amino acids.
  • Challenges in quantifying modified residues can be mitigated with appropriate standards.