Optimizing Sequence Coverage for a Moderate Mass Protein in Nano-Electrospray Ionization Quadrupole Time-of-Flight Mass Spectrometry

Authors

Ryan E. Matsuda, University of Nebraska-LincolnFollow
Venkata Kolli, University of Nebraska-LincolnFollow
Megan Woods, University of Nebraska-Lincoln
Eric D. Dodds, University of Nebraska-LincolnFollow
David S. Hage, University of Nebraska - LincolnFollow

Date of this Version

6-2016

Citation

R. Matsuda, V. Kolli, M. Woods, E.D. Dodds, D.S. Hage, Optimizing Sequence Coverage for a Moderate Mass Protein in Nano-Electrospray Ionization Quadrupole Time-of- Flight Mass Spectrometry, Analytical Biochemistry (2016), doi: 10.1016/j.ab.2016.06.014.

Comments

Copyright 2016 Elsevier. Used by permission.
This is a PDF file of an unedited manuscript that has been accepted for publication.

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 matrixassisted 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.