Deconvolving Native and Intact Protein Mass Spectra with UniDec

Marius M. Kostelic, Michael T. Marty

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations


Intact protein, top-down, and native mass spectrometry (MS) generally requires the deconvolution of electrospray ionization (ESI) mass spectra to assign the mass of components from their charge state distribution. For small, well-resolved proteins, the charge can usually be assigned based on the isotope distribution. However, it can be challenging to determine charge states with larger proteins that lack isotopic resolution, in complex mass spectra with overlapping charge states, and in native spectra that show adduction. To overcome these challenges, UniDec uses Bayesian deconvolution to assign charge states and to create a zero-charge mass distribution. UniDec is fast, user-friendly, and includes a range of advanced tools to assist in intact protein, top-down, and native MS data analysis. This chapter provides a step-by-step protocol and an in-depth explanation of the UniDec algorithm, and highlights the parameters that affect the deconvolution. It also covers advanced data analysis tools, such as macromolecular mass defect analysis and tools for assigning potential PTMs and bound ligands. Overall, this chapter provides users with a deeper understanding of UniDec, which will enhance the quality of deconvolutions and allow for more intricate MS experiments.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages22
StatePublished - 2022

Publication series

NameMethods in Molecular Biology


  • Bayesian deconvolution
  • Electrospray ionization
  • Intact protein analysis
  • Mass spectrometry data analysis
  • Nanodiscs
  • Native mass spectrometry
  • Top-down proteomics

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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