Matrix-assisted Laser Desorption/Ionization Mass Spectrometry Sample Preparation Techniques Designed for Various Peptide and Protein Analytes

Kussmann, Martin; Nordhoff, Eckhard; Rahbek-Nielsen, Henrik; Haebel, Sophie; Rossel-Larsen, Martin; Jakobsen, Lene; Gobom, Johan; Mirgorodskaya, Ekatarina; Kroll-Kristensen, Anne; Palm, Lisbeth; Roepstorff, Peter

doi:10.1002/(sici)1096-9888(199706)32:6<593::aid-jms511>3.0.co;2-d

Cited by 393 publications

(191 citation statements)

References 3 publications

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“…After incubation for 1 h in HFIP, the solvent was evaporated by a stream of nitrogen. The HFIP-treated samples were then redissolved in a minimal volume of 1:1 acetonitrile-H 2 O with 0.1% trifluoroacetic acid (TFA) and applied to the sample plate using the sandwich method with sinapinic acid (Thermo Fisher Scientific) as the MALDI matrix 59 . The dried samples were then analysed on a Bruker ultrafleXtreme MALDI-TOF/TOF in linear mode.…”

Section: And 3)mentioning

confidence: 99%

Programmable biofilm-based materials from engineered curli nanofibres

et al. 2014

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The significant role of biofilms in pathogenicity has spurred research into preventing their formation and promoting their disruption, resulting in overlooked opportunities to develop biofilms as a synthetic biological platform for self-assembling functional materials. Here we present Biofilm-Integrated Nanofiber Display (BIND) as a strategy for the molecular programming of the bacterial extracellular matrix material by genetically appending peptide domains to the amyloid protein CsgA, the dominant proteinaceous component in Escherichia coli biofilms. These engineered CsgA fusion proteins are successfully secreted and extracellularly self-assemble into amyloid nanofibre networks that retain the functions of the displayed peptide domains. We show the use of BIND to confer diverse artificial functions to the biofilm matrix, such as nanoparticle biotemplating, substrate adhesion, covalent immobilization of proteins or a combination thereof. BIND is a versatile nanobiotechnological platform for developing robust materials with programmable functions, demonstrating the potential of utilizing biofilms as large-scale designable biomaterials.

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Section: And 3)mentioning

confidence: 99%

Programmable biofilm-based materials from engineered curli nanofibres

et al. 2014

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“…Native N-glycan samples were additionally desalted on the graphitized carbon column [22] prior to ESI MS. Mini cartridges were prepared by dismantling a normal-sized GlycoClean H cartridge (Oxford GlycoSciences, UK) and packing the material into disposable pipette tips as generally described earlier [23]. Prior to use, the columns were washed with three changes of 50 l 80% acetonitrile (vol/vol) in 0.1% (vol/vol) TFA followed by 3 ϫ 100 l of water.…”

Section: In Vitro ␣1-3fucosylation Of the Trisialylated N-glycansmentioning

confidence: 99%

Sequencing of tri- and tetraantennary N-glycans containing sialic acid by negative mode ESI QTOF tandem MS

Sagi¹,

Peter‐Katalinić²,

Conradt

et al. 2002

J. Am. Soc. Mass Spectrom.

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Application of the negative mode electrospray ionization-quadrupole time-of-flight mass spectrometry (ESI QTOF) tandem MS for determination of substitution patterns by sialic acid and/or fucose and extention by additional LacNAc disaccharide units in single branches of multiantennary N-glycans from biological samples is described. Fragmentation patterns which can be obtained by low energy collision-induced dissociation (CID) using the QTOF instrument include cleavage ions, diagnostic for determination of antennarity and for specific structural features of single antennae. Systematic fragmentation studies in the negative ion mode were focussed toward formation of the D diagnostic ion relevant for assignment of 3-and 6-antennae in complex N-glycans carrying three and four antennae in combination with epitope-relevant B-and C-type ions. For validation of this approach ESI QTOF fragmentation of the permethylated analogues was carried out in the positive ion mode. Using this strategy, products of in vitro glycosylation reactions were investigated in order to clarify some general aspects of N-glycan acceptor specificity during biosynthesis. ␣1-3fucosylation using GDPfucose along with a soluble form of the recombinant human ␣1-3fucosyltransferase VI was carried out on tri-and tetraantennary precursors to test structural requirements for formation of Le A number of biologically relevant glycoproteins contain large N-linked complex carbohydrate moieties, proposed to be involved in specific interactions with other molecules and/or cell surfaces. Highly regulated biosynthetic pathways in which Nacetylglucosaminyltransferases are incorporating GlcNAc residues on the mannotriose core are determining factors for the formation of a number of antennae in complex-type N-glycans [1]. In multiantennary glycans different antennae can show different extent of elongation by N-acetyllactosaminyl repeats and number and site of modifications by sialic acid and fucose moieties, contributing to the microheterogeneity on the single glycosylation site. Besides, the expression of a certain oligosaccharide epitope may be restricted to just one of the antennae present. In order to carry on a complete characterization of complex type N-glycan mixtures obtained from glycoproteins after release by Nglycanase, combined chromatographic, mass spectrometric and nuclear magnetic resonance spectroscopic approach can be mandatory, as demonstrated in the case of human erythropoietin (hEPO), a glycoprotein containing bi-, tri-and tetraantennary N-glycan structures [2]. Its biological activity in terms of the clearance rate was positively correlated with the ratio of tetra-to biantennary N-glycans, where the hEPO with wellbranched tetraantennary glycans remained at higher levels in the plasma [3]. Particular attention is to be paid to identification of glycosylation patterns in terms of antennarity in glycoproteins from different organs, since they may express identical protein cores, carrying glycoforms of different antennarity and therefore different ant...

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“…M atrix assisted laser ionization/desorption mass spectrometry (MALDI-MS) is one of the most powerful tools for analysis of large biomolecules, such as peptides, proteins, and nucleic acids [1][2][3][4][5][6][7]. Typical sample preparation involves mixing the analyte with organic UV-absorbing matrices.…”

mentioning

confidence: 99%

CHCA-modified Au nanoparticles for laser desorption ionization mass spectrometric analysis of peptides

Duan

Linman

Chen

et al. 2009

J. Am. Soc. Mass Spectrom.

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Gold nanoparticles (AuNPs) have been studied as a potential solid-state matrix for laser desorption/ionization mass spectrometry (LDI-MS) but the efficiency in ionization remains low. In this report, AuNPs are capped by a self-assembled monolayer of cysteamine and modified with ␣-cyano-4-hydroxycinnanic acid (CHCA) for effective MALDI measurements. CHCA-terminated AuNPs offer marked improvement on peptide ionization compared with citrate-capped or cysteamine-capped AuNPs. The coating also effectively suppresses formation of Au cluster ions and analyte fragment ions, leading to cleaner mass spectra. Addition of glycerol and citric acid to the peptide/AuNPs sample further improves the performance of these AuNPs for LDI-MS analysis. Glycerol appears to enhance the dispersion of AuNPs in sample spots, increasing the sample ionization and shot-to-shot reproducibility, while citric acid serves as an external proton donor, providing high production of protonated analyte ions and reducing fragmentation of peptides on the nanoparticle-based surface. Optimal ratios of citric acid, glycerol, and AuNPs in sample solution have been systematically studied. A more than 10-fold increase for desorption ionization of peptides can be achieved by combining 5% glycerol and 20 mM citric acid with the CHCA-terminated AuNPs. The applicability of the CHCA-AuNPs for LDI-MS analysis of protein digests has also been demonstrated. This work shows the potential of AuNPs for SALDI-MS analysis, and the improvement with chemical functionalization, controlled dispersion, and use of an effective proton donor. (J Am Soc Mass

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Matrix-assisted Laser Desorption/Ionization Mass Spectrometry Sample Preparation Techniques Designed for Various Peptide and Protein Analytes

Cited by 393 publications

References 3 publications

Programmable biofilm-based materials from engineered curli nanofibres

Programmable biofilm-based materials from engineered curli nanofibres

Sequencing of tri- and tetraantennary N-glycans containing sialic acid by negative mode ESI QTOF tandem MS

CHCA-modified Au nanoparticles for laser desorption ionization mass spectrometric analysis of peptides

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