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Related Experiment Video

Updated: Oct 17, 2025

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tipNrich: A Tip-Based N-Terminal Proteome Enrichment Method.

Seonjeong Lee1,2, Shinyeong Ju1, Seok Jin Kim3,4

  • 1Center for Theragnosis, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea.

Analytical Chemistry
|October 7, 2021
PubMed
Summary

We developed tipNrich, a novel method for analyzing N-terminal protein modifications from scarce samples. This technique enables sensitive N-terminome profiling for disease diagnosis and protein quality control.

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Area of Science:

  • Proteomics
  • Biochemistry
  • Analytical Chemistry

Background:

  • Mass spectrometry analysis of protein post-translational modifications typically requires substantial sample amounts.
  • Analyzing limited protein samples, such as clinical biopsies, presents significant challenges for current methods.

Purpose of the Study:

  • To introduce tipNrich, a highly optimized, tip-based method for N-terminal protein analysis.
  • To enable sensitive identification of N-terminal peptides from femtomole-scale protein quantities.
  • To demonstrate the utility of tipNrich in disease diagnosis and protein quality assessment.

Main Methods:

  • A novel, single-pipette-tip procedure (tipNrich) was developed to minimize sample loss.
  • Low-resolution mass spectrometry was used to investigate purified single proteins from various organisms.
  • Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was integrated to accelerate analysis time.

Main Results:

  • tipNrich successfully identified N-terminal peptides with various modifications, including ragged N-termini, from limited protein amounts.
  • The method demonstrated high sensitivity, analyzing even femtomole-scale quantities of single proteins.
  • N-terminome profiling using tipNrich differentiated wild-type transthyretin amyloidosis patients from healthy individuals.

Conclusions:

  • tipNrich significantly enhances the ability to identify N-terminal peptides from extremely scarce protein samples.
  • The method offers faster processing times, beneficial for protein product quality checks.
  • tipNrich is a valuable tool for characterizing diseases related to N-terminal proteoforms and for clinical diagnostics.