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Best practices and benchmarks for intact protein analysis for top-down mass spectrometry.

  • Author(s): Donnelly, Daniel P
  • Rawlins, Catherine M
  • DeHart, Caroline J
  • Fornelli, Luca
  • Schachner, Luis F
  • Lin, Ziqing
  • Lippens, Jennifer L
  • Aluri, Krishna C
  • Sarin, Richa
  • Chen, Bifan
  • Lantz, Carter
  • Jung, Wonhyeuk
  • Johnson, Kendall R
  • Koller, Antonius
  • Wolff, Jeremy J
  • Campuzano, Iain DG
  • Auclair, Jared R
  • Ivanov, Alexander R
  • Whitelegge, Julian P
  • Paša-Tolić, Ljiljana
  • Chamot-Rooke, Julia
  • Danis, Paul O
  • Smith, Lloyd M
  • Tsybin, Yury O
  • Loo, Joseph A
  • Ge, Ying
  • Kelleher, Neil L
  • Agar, Jeffrey N
  • et al.
Abstract

One gene can give rise to many functionally distinct proteoforms, each of which has a characteristic molecular mass. Top-down mass spectrometry enables the analysis of intact proteins and proteoforms. Here members of the Consortium for Top-Down Proteomics provide a decision tree that guides researchers to robust protocols for mass analysis of intact proteins (antibodies, membrane proteins and others) from mixtures of varying complexity. We also present cross-platform analytical benchmarks using a protein standard sample, to allow users to gauge their proficiency.

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