A new method to improve sensitivity and resolution in matrix-assisted laser desorption/ionization time of flight mass spectrometry.
High detection sensitivity and resolution are two critical parameters for recording good peptide mass fingerprints (PMF) of low abundance proteins. This paper reports a mass spectrometry (MS) sample preparation technique that could improve sensitivity and resolution. By coating the MS steel target with a thin layer of pentadecafluorooctamido propyltrimethoxysilane, which was both polar and nonpolar solvent repellent, the transferred sample droplets on its surface were significantly smaller. As a result, the analyte of the peptide mixture became more concentrated and homogeneous, which helped to improve the sensitivity. The advantages of a modified MS target were documented by mass spectra improvement of attomole level standard peptides and silver-stained proteins from polyacrylamide gels. The mass signal of angiotensin II at 100 attomole was difficult to record on the conventional support, whereas it was easily detected on the modified one. The PMF of cytochrome C was also better recorded on the modified support, in terms of both signal-to-noise ratio and the number of detected peptides. When silver-stained proteins from two-dimensional electrophoresis gels were analyzed, in most cases more satisfactory peptide mass spectra were obtained from the modified support. Searching protein databases with more mass data from the improved PMFs, several unknown proteins were successfully identified.
Xiong
Shaoxiang
S
Beijing Mass Spectrometry Center, Center for Molecular Science, Institute of Chemistry, Beijing, PR China.
Ding
Qinxue
Q
Zhao
Zhengwen
Z
Chen
Wenzhang
W
Wang
Guanghui
G
Liu
Shaojun
S
D000804:Angiotensin II / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D045304:Cytochromes c / Q000737:chemistry; D019992:Databases as Topic; D004591:Electrophoresis, Polyacrylamide Gel; D011138:Polytetrafluoroethylene / Q000737:chemistry; D012680:Sensitivity and Specificity; D012821:Silanes / Q000737:chemistry; D016622:Silver Staining; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D012821:Silanes; D000804:Angiotensin II; D011138:Polytetrafluoroethylene; D045304:Cytochromes c