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

Fingerprint

Electrospray ionization
Mass spectrometry
Mass Spectrometry
Peptides
Proteins
Fractionation
Serum Albumin
Trypsin
Ionization
Desorption
Lasers

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

Cite this

Optimizing sequence coverage for a moderate mass protein in nano-electrospray ionization quadrupole time-of-flight mass spectrometry. / Matsuda, Ryan; Kolli, Venkata; Woods, Megan; Dodds, Eric D; Hage, David S.

In: Analytical Biochemistry, Vol. 509, 15.09.2016, p. 115-117.

Research output: Contribution to journalArticle

@article{e757ee2b38264be3afbd2522906410d0,
title = "Optimizing sequence coverage for a moderate mass protein in nano-electrospray ionization quadrupole time-of-flight mass spectrometry",
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.",
keywords = "Collision-induced dissociation, Human serum albumin, Internal calibration, Nano-electrospray ionization quadrupole time-of-flight mass spectrometry, Sample pretreatment, Sequence coverage",
author = "Ryan Matsuda and Venkata Kolli and Megan Woods and Dodds, {Eric D} and Hage, {David S}",
year = "2016",
month = "9",
day = "15",
doi = "10.1016/j.ab.2016.06.014",
language = "English (US)",
volume = "509",
pages = "115--117",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Optimizing sequence coverage for a moderate mass protein in nano-electrospray ionization quadrupole time-of-flight mass spectrometry

AU - Matsuda, Ryan

AU - Kolli, Venkata

AU - Woods, Megan

AU - Dodds, Eric D

AU - Hage, David S

PY - 2016/9/15

Y1 - 2016/9/15

N2 - 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.

AB - 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.

KW - Collision-induced dissociation

KW - Human serum albumin

KW - Internal calibration

KW - Nano-electrospray ionization quadrupole time-of-flight mass spectrometry

KW - Sample pretreatment

KW - Sequence coverage

UR - http://www.scopus.com/inward/record.url?scp=84978427775&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84978427775&partnerID=8YFLogxK

U2 - 10.1016/j.ab.2016.06.014

DO - 10.1016/j.ab.2016.06.014

M3 - Article

VL - 509

SP - 115

EP - 117

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

ER -