Date of this Version
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.
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.