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 language | English (US) |
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Pages (from-to) | 115-117 |
Number of pages | 3 |
Journal | Analytical Biochemistry |
Volume | 509 |
DOIs | |
State | Published - Sep 15 2016 |
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