Optimizing sequence coverage for a moderate mass protein in nano-electrospray ionization quadrupole time-of-flight mass spectrometry

Ryan Matsuda, Venkata Kolli, Megan Woods, Eric D. Dodds, David S. Hage

Research output: Contribution to journalArticle

Abstract

Sample pretreatment was optimized to obtain high sequence coverage for human serum albumin (HSA, 66.5 kDa) when using nano-electrospray ionization quadrupole time-of-flight mass spectrometry (nESI–Q-TOF–MS). Use of the final method with trypsin, Lys-C, and Glu-C digests gave a combined coverage of 98.8%. The addition of peptide fractionation resulted in 99.7% coverage. These results were comparable to those obtained previously with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI–TOF–MS). The sample pretreatment/nESI–Q-TOF–MS method was also used with collision-induced dissociation to analyze HSA digests and to identify peptides that could be employed as internal mass calibrants in future studies of modifications to HSA.

Original languageEnglish (US)
Pages (from-to)115-117
Number of pages3
JournalAnalytical Biochemistry
Volume509
DOIs
StatePublished - Sep 15 2016

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Keywords

  • Collision-induced dissociation
  • Human serum albumin
  • Internal calibration
  • Nano-electrospray ionization quadrupole time-of-flight mass spectrometry
  • Sample pretreatment
  • Sequence coverage

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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