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Automated Acquisition and Analysis of Data for Monitoring Protein Conjugation by LC–MS Using BioPharma Compass
Malcolm J. Saxton, Ryan Hylands, Karl Nichols, Joanne Waters, Stuart Pengelley, and Wolfgang Jabs, Bruker Daltonics
In the example reported here, albumin was conjugated with a fluorescent dye. The success of this experiment was judged according to the percentage of correct conjugation state that was detected at different time points during a time course experiment, with the correct conjugation state assigned as being a 1:1 ratio of albumin:dye. The intact mass of all protein conjugates produced was calculated and used to determine the conjugation ratios for the various conjugation states that were produced at a given time point, based on signal intensity. From this, the percentage of protein that was detected in the correct conjugation state could be used as an indication of the purity of the final conjugated product.
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Combining PGC-LC ESI and MS2-MS3 Ion Trap Analysis for Structural Characterization of Sulphated O-glycans
Kathrin Stavenhagen, Daniel Kolarich, Glycoproteomics Group, Department of Biomolecular Systems; Max-Planck Institue of Colloids and Interfaces; and Ulrike Schweiger-Hufnagel, Kristina Marx, and Markus Meyer, Bruker Daltonics
Porous graphitized carbon (PGC) LC-ESI ion trap MSn analysis provides an attractive alternative to elucidate fine structural details that otherwise are difficult if not impossible to differentiate from the limited amount of biological sample material. Using human saliva as an easily available source of mucin type O-glycans, the advantages of ion trap MSn in combination with PGC separation for detailed structure determination is exemplified on a sulphated tetrasaccharide O-glycan detected in four different isoforms.
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Smart MRM with Electron Transfer Dissociation of Hemoglobin in Clinical Samples
D. Coelho Graça, Pierre Lescuyer, Lorella Clerici, Yury O. Tsybin, Kaveh Samii, Denis Hochstrasser, Alexander Scherl, Ralf Hartmer, and Markus Meyer Bruker Daltonik GmbH
Today, biological diagnosis of hemoglobinopathies requires the combination of different measurements and instrumentations. The current complex procedure generally requires hematological tests, liquid chromatography, capillary electrophoresis, or electrophoresis, with results being confirmed by molecular biology.
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High Throughput Quantitative and Qualitative DMPK Measurements
Dr. Don Richards, Bruker Daltonics and Helen Lomax, Fortis Technologies
This application note explains the use of quan/qual tools with fast chromatography suitable for high throughput screen of P450 microsomal metabolism of drugs in a DMPK arena.
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Food Authenticity – Classification of Coffee Types Based on LC-MS
Verena Tellström, Sven Brand, Michael Becker, Klaus Meyer, Gabriela Zurek, and Aiko Barsch, Bruker Daltonics
In a recent study, the compact QTOF System was used to analyze extracts from 13 different types of coffee capsule. A non-targeted metabolomics workflow enabled differentiation of coffee types based on their flavor intensity and readily identified the compounds responsible for the differentiation. In this study, which serves as an example for food authenticity assessment in general, the established statistical PCA and PLS models were used to classify different coffee samples into similar or distinct groups, and to predict the relative intensity for each group.
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Plant Metabolomics Research Highlight: Prof. Lloyd W. Sumner Combines UHPLC-MS-SPE-NMR and Prof. Kazuki Saito Employs FT-ICR-MS in Novel Strategies to Identify Unknown Plant Metabolites
Lloyd W. Sumner, Zhentian Lei, Dennis Fine, Daniel Wherritt, David V. Huhman, Wiebke Timm, Gabriela Zurek, Aiko Barsch, Kota Kera, Hidezuki Suzuki, Ryo Nakabayashi, and Kazuki Saito, Bruker Daltonics
Strategies based on hyphenated techniques such as LC-MS-SPE-NMR and FT-ICR-MS enable high-throughput structure elucidation which will pave the way to identifying the vast number of yet unknownplant secondary metabolites, and ultimately complete the picture of entire plant metabolomes. In this application note we highlight workflows recently implemented in professors Sumner's and Saito's laboratories addressing the need to identify unknown compounds in plant metabolomics.
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MALDI Imaging with Single Cell Resolution at 10 μm Pixel Size
Michael Becker, Eckhard Belau, Sören-Oliver Deininger, Jens Fuchser, and Shannon Cornett, Bruker Daltonics
Key parameters for MALDI imaging are spatial resolution and mass spectral quality. In this application note, we explore the feasibility of single cell imaging. Typically, high spatial resolution requires matrix preparation that is drier than usual and often is limited to high abundant lipid species. For the high resolution imaging presented here, we utilized a dry sublimation device, which enabled us to perform MALDI imaging at a spatial resolution of 10 μm and to resolve individual cells in tissue images. The 10 μm pixel size was achieved on both solarix FTMS and autofleXspeed and ultrafleXtreme platforms.
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Definitive Elemental Formula Determination Debuts with SmartFormula-HR™
Christopher Thompson, Michael Easterling, and Darwin Asa, Bruker Daltonics
In this study, SmartFormula-HR Software is applied to a mixture of pesticides. The mixture is first separated via HPLC, and then detected with high resolution mass spectrometry using the solariX.
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Top Down Analysis of Histone H4
Jeremy J. Wolff and Christopher J. Thompson, Bruker Daltonics
Characterization of proteins by top-down mass spectrometry is often necessary when many post translational modifications are present. Histones are an example of this type of challenge due to their abundant and varying modifications. We demonstrate top-down dissociation of two intact human histone H4 proteins with ECD, ETD, CID, and MALDI-ISD on the Bruker solariX FTMS platform. These dissociation techniques were used to determine the presence of both acetylation and dimethylation on the proteins.
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Microbial Imaging Mass Spectrometry with Fourier Transform Ionization Mass Spectrometry
Vanessa Phelan, Peiter C. Dorrestein, and Shannon Cornett, Bruker Daltonics
In this application note we describe a straightforward approach for the preparation and molecular analysis of a bacterial-fungal interaction and the visualization of metabolic exchange factors using an FT-ICR high resolution mass spectrometer.
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