title: Liquid MALDI MS Analysis of Complex Peptide and Proteome Samples.
abstract: Matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) is well-known to be a powerful technique for the analysis of biological samples. By using glycerol-based liquid support matrices (LSMs) instead of conventional MALDI matrices the power of this technique can be extended further. In this study, we exploited LSMs for the identification of complex samples, that is, the Lactobacillus proteome and a bovine serum albumin (BSA) digest. Liquid and solid MALDI samples were manually and robotically prepared by coupling a nanoflow high-performance liquid chromatography (nanoHPLC) system to an automated MALDI sample spotting device. MS and MS/MS data were successfully acquired at the femtomole level using TOF/TOF as well as Q-TOF instrumentation and used for protein identification searching sequence databases. For the BSA digest analysis, liquid MALDI samples resulted in peptide mass fingerprints, which led to a higher confidence in protein identification compared with solid (crystalline) MALDI samples; however, postsource decay (PSD) MS/MS analysis of both the proteome of Lactobacillus plantarum WCFS1 cells and BSA digest showed that further optimization of the formation and detection of peptide fragment ions is still needed for liquid MALDI samples, as the MS/MS ion search score was lower than that for the solid MALDI samples, reflecting the poorer quality of the liquid MALDI-PSD spectra, which can be attributed to the differences in PSD parameters and their optimization that is currently achievable.
authors.affiliation: Department of Chemistry, University of Reading , Whiteknights, Reading, RG6 6AD, United Kingdom.
authors.forename: Kanjana
authors.initials: K
authors.lastname: Wiangnon
authors.affiliation: Department of Chemistry, University of Reading , Whiteknights, Reading, RG6 6AD, United Kingdom.
authors.forename: Rainer
authors.initials: R
authors.lastname: Cramer
mesh_terms: D000818:Animals; D002417:Cattle; D005990:Glycerol; D007778:Lactobacillus / Q000737:chemistry; D010449:Peptide Mapping; D010455:Peptides / Q000032:analysis*; D020543:Proteome / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D010455:Peptides; D020543:Proteome; D012710:Serum Albumin, Bovine; D005990:Glycerol
keywords: LC−MALDI; MALDI MS/MS; MALDI-CID; MALDI-PSD; MALDI-TOF; Q-TOF; automation; liquid MALDI; liquid support matrix; mass spectrometry