MALDI In-Source Decay, from sequencing to imaging

in Topics in Current Chemistry (in press)

MALDI is now a mature method allowing the identification and, more challenging, the quantification of biopolymers (proteins, nucleic acids, glycans…). MALDI spectra show mostly intact singly charged ions ... [more ▼]

MALDI is now a mature method allowing the identification and, more challenging, the quantification of biopolymers (proteins, nucleic acids, glycans…). MALDI spectra show mostly intact singly charged ions. To obtain fragments, the activation of singly charged precursors is necessary, but not efficient above 3.5 kDa thus making MALDI MS/MS difficult for large species. In-source decay (ISD) is a prompt fragmentation reaction that can be induced thermally or by radicals. As fragments are formed in the source, precursor ions cannot be selected; however, the technique is not limited by the mass of the analyzed compounds and pseudo MS/MS can be performed on intense fragments. The discovery of new matrices that enhance the ISD yield, combined with the high sensitivity of MALDI mass spectrometers, and software development, opens new perspectives. We first review the mechanisms involved in the ISD processes, then discuss ISD applications like top-down sequencing and post-translational modifications studies, and finally review MALDI-ISD tissue imaging applications. [less ▲]

Determination of chloramphenicol in honey, 1 shrimp and poultry meat with liquid chromatography-mass spectrometry. Validation of the method according to Commission Decision 2002/657/EC

in Food Analytical Methods (in press)

Chloramphenicol (CAP) is an antibiotic used for the treatment of bacterial infections in human and veterinary medicine. The use of CAP was prohibited in the European Union in 1994. Control laboratories ... [more ▼]

Chloramphenicol (CAP) is an antibiotic used for the treatment of bacterial infections in human and veterinary medicine. The use of CAP was prohibited in the European Union in 1994. Control laboratories are required to use suitably validated analytical methods to check sample compliance with the regulation. A quantitative method based on liquid chromatography coupled to isotopic dilution tandem mass spectrometry (LC–IDMS/MS) was developed for the determination of chloramphenicol in honey, shrimp and poultry meat. The experimental protocol consisted of a liquid-liquid extraction with ethyl acetate. Separation and detection were realized respectively by a 2690 Waters HPLC (Milford, MA, USA) and a Micromass Triple Quadrupole mass spectrometer (Micromass, Manchester, UK), equipped with an Electrospray source. The effects of mobile phase additives on the response of LC/ESI/MS were examined. Two different HPLC columns were tested: the X-Terra from Waters and the Alltima HP C18 HL from Alltech (Deerfield, IL, USA). A validation of the method was conducted according to the EU criteria for the analysis of chloramphenicol in foods. The decision limits (CCα) were 0.04, 0.03, 0.07 µg kg-1 and the detection capabilities (CCβ) were 0.05, 0.04, 0.08 µg kg-1 for honey, shrimp and poultry meat respectively. Those values are below the minimum required performance limit (MRPL) set at 0.3 µg kg-1 by the EU and 0.1 µg kg-1 by Belgium. Our protocol has the advantage to propose a unique extraction method working as well for honey, shrimp and poultry meat, contrary to similar published methods in which a different extraction method is used for each type of matrix. [less ▲]

Les venins d'animaux, nouvelle panacée?

in Athena (2013)

Araignées, serpents, scorpions,… autant d’animaux ayant une place particulière dans l’imaginaire collectif. Fascinants, horripilants voire même terrifiants, les adjectifs ne manquent pas pour qualifier ... [more ▼]

Araignées, serpents, scorpions,… autant d’animaux ayant une place particulière dans l’imaginaire collectif. Fascinants, horripilants voire même terrifiants, les adjectifs ne manquent pas pour qualifier les réactions qu’ils suscitent auprès des populations. Que dire alors de la peur engendrée par leur venin. Une simple piqûre, morsure ou contact peut s’avérer extrêmement dangereux voire létal... [less ▲]

Ultraviolet Laser Induced Hydrogen Transfer Reaction: Study of the First Step of MALDI In-Source Decay Mass Spectrometry

in Journal of Physical Chemistry B (2013), 117(8), 2321-2327

The early mechanisms of matrix-assisted laser desorption/ionization in-source decay (MALDI-ISD) are described herein. MALDI-ISD is initiated by the hydrogen transfer from excited matrix molecules to the ... [more ▼]

The early mechanisms of matrix-assisted laser desorption/ionization in-source decay (MALDI-ISD) are described herein. MALDI-ISD is initiated by the hydrogen transfer from excited matrix molecules to the carbonyl oxygen of the peptide backbone, which is followed by a radical-induced cleavage, producing the c′/z• fragment pair. As expected, the use of 2,5-DHB or 1,5-DAN was efficient to induce MALDI-ISD, and the strongest intensity of MALDI-ISD fragments was observed when laser shots were performed on matrix crystals. In contrast, the hydrogen radical transfer reaction was suppressed by using ionic liquid and amorphous structure of 2,5-DHB and 1,5-DAN mixture as a matrix. Our results suggest that the hydrogen transfer occurs on the matrix crystal during the dissipation of the laser energy and before desorption, following ISD fragments formed in the MALDI plume. [less ▲]

Selected Protein Monitoring in Histological Sections by Targeted MALDI-FTICR in-source decay Imaging.

in Analytical Chemistry (2013), sous presse

MALDI mass spectrometry imaging (MALDI MSI) is a rapidly growing method in biomedical research allowing molecular mapping of proteins on histological sections. The images can be analyzed in terms of ... [more ▼]

MALDI mass spectrometry imaging (MALDI MSI) is a rapidly growing method in biomedical research allowing molecular mapping of proteins on histological sections. The images can be analyzed in terms of spectral pattern to define regions of interest. However, the identification and the differential quantitative analysis of proteins require off line or in situ proteomic methods using enzymatic digestion. The rapid identification of biomarkers holds great promise for diagnostic research but the major obstacle is the absence of rapid and direct method to detect and identify with a sufficient dynamic range a set of specific biomarkers. In the current work, we present a proof of concept for a method allowing identifying simultaneously a set of selected biomarkers on histological slices with minimal sample treatment using in-source decay (ISD) MSI and MALDI-Fourier transform ion cyclotron resonance (FTICR). In the proposed method, known biomarkers are spotted next to the tissue of interest, the whole MALDI plate being coated with 1,5-DAN matrix. The latter enhances MALDI radical-induced ISD, providing large tags of the amino acid sequences. Comparative analysis of ISD fragments between the reference spots and the specimen in imaging mode allows for unambiguous identification of the selected biomarker while preserving full spatial resolution. Moreover, the high resolution/high mass accuracy provided by FTICR mass spectrometry allows the identification of proteins. Well-resolved peaks and precise measurements of masses and mass differences allow the construction of reliable sequence tags for proteins identification. The method will allow the use MALDI-FTICR MSI as method for rapid targeted biomarker detection in complement to classical histology. [less ▲]

Proteomic investigation of aphid honeydew reveals an unexpected diversity of proteins

in PLoS ONE (2013), Accepted pending revision

Preliminary assessment of the risk linked to furan ingestion by babies consuming only ready-to-eat food

in Food Additives & Contaminants. Part A. Chemistry, Analysis, Control, Exposure & Risk Assessment (2013)

The risk linked to furan ingestion has been assessed in previous papers for Belgian adults and children (Scholl et al., 2012b; Scholl et al., 2012c). The present paper focuses on infants consuming only ... [more ▼]

The risk linked to furan ingestion has been assessed in previous papers for Belgian adults and children (Scholl et al., 2012b; Scholl et al., 2012c). The present paper focuses on infants consuming only ready-to-eat baby food. As there is no Belgian baby dietary database, the furan exposure assessment was carried out by using Italian infant consumption database and Belgian contamination data. The estimated daily intake (EDI) was calculated according to a deterministic methodology. It involved 42 commercially available ready-to-eat baby food and 36 baby consumption records. The mean EDI was 1,460 ng * (kgb.w.*day)-1 which is 3.8 times higher than the 381 ng * (kgb.w.*day)-1 reported for Belgian adults, and 3.5 times higher than the 419 ng * (kgb.w. * day)-1 measured for Belgian children. To assess and characterize the risk for babies exposure the Margin of Exposure (MoE) was calculated. It highlighted that 74% of infants have a MoE below 1,000, with a minimum of 140. However, these are only preliminary results as they were calculated from a very small dataset and the infant cytochrome P450 activity is significantly different compared to the adult. Therefore, the risk linked to furan ingestion by babies should be assessed in a different manner. To this end, additional data regarding a baby diet as well as a better understanding of furan toxicity for babies are needed to characterize more accurately the risk for infants. [less ▲]

Ion-Mobility mass spectrometry as a potential tool to assign disulfide bonds arrangements in peptides with multiple disulfide bridges.

in Analytical Chemistry (2013)

Disulfide bridges play a major role in defining the structural properties of peptides and proteins. However, the determination of the cysteine pairing is still challenging. Peptide sequences are usually ... [more ▼]

Disulfide bridges play a major role in defining the structural properties of peptides and proteins. However, the determination of the cysteine pairing is still challenging. Peptide sequences are usually achieved using MS/MS spectra of the totally reduced unfolded species but the cysteine pairing information is lost. On the other hand, MS/MS experiments performed on native folded species show complex spectra composed of non-classical ions. MS/MS alone does not allow the cysteine pairing nor the full sequence of an unknown peptide to be determined. The major goal of this work is to set up a strategy for the full structural characterization of peptides including disulfide bridges annotation in the sequence. This strategy was developed by combining Ion Mobility Spectrometry (IMS)and Collision Induced Dissociation(CID). It is assumed that the opening of one S-S bridges in a peptide leads to a structural evolution which results in a modification of IMS drift time. In the presence of multiple S-S bridges, the shift in arrival time will depend on which disulfide(s) has (have) been reduced and on the shape adopted by the generated species. Due to specific fragmentations observed for each species, CID experiments performed after the mobility separation could provide not only information on peptide sequence, but also on the localization of the disulfide bridges. To achieve this goal, synthetic peptides containing two disulfides were studied. The openings of the bridges were carried out following different experimental conditions such as reduction, reduction/alkylation or oxidation. Due to disulfide scrambling highlighted with the reduction approaches, oxidation of S-S bonds into cysteic acids appeared to be the best strategy. Cysteines connectivity was then unambiguously determined for the two peptides, without any disulfide scrambling interference. [less ▲]

Secretion and maturation of conotoxins in the venom ducts of Conus textile

in Toxicon (2012), 60(8), 1370-1379

Utilisation des termites comme source de microorganismes dans la filière de production du bioéthanol de seconde génération

Poster (2012, November 14)

Les termites abritent une microflore symbiotique qui intervient dans la dégradation des fibres constitutives du bois, synthétisant des enzymes capables d’hydrolyser ses composants. Les sucres ... [more ▼]

Les termites abritent une microflore symbiotique qui intervient dans la dégradation des fibres constitutives du bois, synthétisant des enzymes capables d’hydrolyser ses composants. Les sucres fermentescibles libérés suite à cette hydrolyse sont utilisables dans le cadre de la production du bioéthanol de seconde génération. [less ▲]