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

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• 作者：Ryan Matsuda ; Venkata Kolli ; Megan Woods ; Eric D. Dodds ; David S. Hage ; ^{dhage1@unl.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Human serum albumin ; Sequence coverage ; Sample pretreatment ; Nano-electrospray ionization quadrupole time-of-flight mass spectrometry ; Collision-induced dissociation ; Internal calibration
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：15 September 2016
• 年：2016
• 卷：509
• 期：Complete
• 页码：115-117
• 全文大小：457 K

文摘

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.