Isomer Separation of Positively Labeled N-glycans by CE-ESI-MS
|
|
- George Garrett
- 5 years ago
- Views:
Transcription
1 Isomer Separation of Positively Labeled N-glycans by CE-ESI-MS G.S.M. Kammeijer Center for Proteomics and Metabolomics CE IN THE BIOTECHNOLOGY & PHARMACEUTICAL INDUSTRIES 19 TH SYMPOSIUM ON THE PRACTICAL APPLICATIONS FOR THE ANALYSIS OF PROTEINS, NUCLEOTIDES AND SMALL MOLECULES (CE PHARM 017) LATE BREAKING SESSION 18 TH OF SEPTEMBER 017
2 GLYCOSYLATION Highly prevalent co- and posttranslational protein modification (PTM) Thought to be the most complex PTM More than 50% of human proteins are glycosylated Large influence on protein function Associated with physiological conditions and the effect of biopharmaceuticals IgG1 [P01857] G.S.M. Kammeijer - g.s.m.kammeijer@lumc.nl
3 GLYCOPROTEOMICS The most abundant monosaccharides in humans are: Glucose Da Galactose Da Mannose Da Fucose Da N-Acetyl-Dglucosamine Da N-Acetyl-Dgalactosamine Da N-Acetylneuraminic acid Da 3 G.S.M. Kammeijer - g.s.m.kammeijer@lumc.nl
4 ANALYSIS APPROACHES INTACT GLYCOPROTEINS GLYCOPEPTIDES RELEASED GLYCANS Different isoforms Protein specific Site-specific Protein specific Analysis of complex samples Not site-specific Requires isolated proteins Fast increase of complexity of the data Not site-specific Not protein-specific 4 G.S.M. Kammeijer - g.s.m.kammeijer@lumc.nl
5 OUTLINE GLYCAN CHARACTERIZATION IMPROVING SENSITIVITY? IMPROVING SEPARATION 5 G.S.M. Kammeijer - g.s.m.kammeijer@lumc.nl
6 IN-HOUSE METHODS GLYCAN ANALYSIS CE-ESI-MS Sample PNGase F release Sample PNGase F release Di-methyl amidation Labeling CE-ESI-MS 6 G.S.M. Kammeijer - g.s.m.kammeijer@lumc.nl
7 TPNG PROFILE WITH CE-ESI-MS BARE-FUSED SILICA CAPILLARY Intens. x Time [min] Intens. x10 5 Injection: 5 psi 60sec 40 nl Time [min] BFS capillary 90 cm, 30µm id, BGE composition: 10 % AA, capillary coolant: 0 C, voltage: 0 kv, injection sample: 5 psi, 60 sec (40 nl) 7 Kammeijer, G.S.M. et al. Manuscript in preparation
8 TPNG PROFILE WITH CE-ESI-MS BARE-FUSED SILICA CAPILLARY VS VS Intens. x10 5 Injection: 5 psi 60sec 40 nl Time [min] BFS capillary 90 cm, 30µm id, BGE composition: 10 % AA, capillary coolant: 0 C, voltage: 0 kv, injection sample: 5 psi, 60 sec (40 nl) 8 Kammeijer, G.S.M. et al. Manuscript in preparation
9 TPNG PROFILE WITH CE-ESI-MS BFS VS DYNAMIC COATED NEUTRAL CAPILLARY Intens. x Bare-Fused Silica Capillary Injection: 5 psi 60sec (40 nl) Time [min] BFS capillary 90 cm, 30µm id,, BGE: 10 % AA, capillary coolant: 0 C, voltage: 0 kv, injection sample: 5 psi, 60 sec (40 nl) Intens. x Dynamic Coated Neutral Capillary Injection: 5 psi 60sec (40 nl) Time [min] Dynamic neutral coated BFS capillary 90 cm, 30µm id,, BGE: 10 % AA, capillary coolant: 0 C, voltage: 0 kv, injection sample: 5 psi, 60 sec (40 nl) 9 Kammeijer, G.S.M. et al. Manuscript in preparation
10 IN SOURCE DECAY Intens. x10 5 Advantage of using CE for glycan characterization In-source decay can be easily detected and corrected Time [min] 10 G.S.M. Kammeijer - g.s.m.kammeijer@lumc.nl
11 IN-SOURCE DECAY Advantage of using CE for glycan characterization In-source decay can be easily detected and corrected Derivatization method 1 Derivatization method Courtesy of Sander Wagt 11 Kammeijer, G.S.M. et al. Manuscript in preparation
12 DATA AUTOMATION LACYTOOLS Jansen, B.C. et al. J. Proteome Res., 016, 15 (7), pp Kammeijer, G.S.M. et al. Manuscript in preparation
13 DATA AUTOMATION ALIGNMENT 13 Kammeijer, G.S.M. et al. Manuscript in preparation
14 DATA AUTOMATION ALIGNMENT Alignment of reference file (Run#1) Alignment of analysis (Run #) Alignment of analysis (Run #3) 14 Kammeijer, G.S.M. et al. Manuscript in preparation
15 DATA AUTOMATION DATA INTEGRATION PEP PEP PEP PEP Aligned mzxml file will be used for further data integration Reference file Provide composition list Expected migration time Migration window Min. charge state # Peak RT Mass Window Time Window Min Charge Max Calibrant protonloss1gip1h5n protonloss1gip1h3n4f protonloss1gip1h4n protonloss1gip1h4n4f protonloss1gip1h4n5f protonloss1gip1h5n4f protonloss1gip1h5n4e protonloss1gip1h5n4f1e protonloss1gip1h5n4e protonloss1gip1h5n4am1e protonloss1gip1h5n4f1e protonloss1gip1h5n5f1e protonloss1gip1h6n5am1e protonloss1gip1h7n6am3e Kammeijer, G.S.M. et al. Manuscript in preparation
16 OUTLINE GLYCAN CHARACTERIZATION IMPROVING SENSITIVITY? IMPROVING SEPARATION 16 G.S.M. Kammeijer - g.s.m.kammeijer@lumc.nl
17 INCREASING SENSITIVITY OF GLYCAN ANALYSIS WITH CE-ESI-MS 17 Significant increase in sensitivity with nano-lc-ms in combination with acetonitrile enriched nitrogen (DEN)- gas ` Nitrogen gas coming in Nitrogen gas passes the headspace of the acetonitrile bottle Dopant enriched nitrogen gas is introduced in the ESI source Acetonitrile in bottle The solvent vapor acts as a dopant for enrichment. The charge state of multiple charged ions are modified optimizing the signal intensity. Improvement factor of approximately one order of magnitude Application Note # LCMS-93 amazon speed ETD: Exploring glycopeptides in protein mixtures using Fragment Triggered ETD and CaptiveSpray nanobooster Kammeijer, G.S.M. et al. Anal. Chem., 016, 88 (11), pp
18 INCREASING SENSITIVITY OF GLYCAN ANALYSIS WITH CE-ESI-MS Applicable on CE-ESI-MS? 18 Kammeijer, G.S.M. et al. Anal. Chem., 016, 88 (11), pp
19 INCREASING SENSITIVITY OF GLYCAN ANALYSIS WITH CE-ESI-MS Background MS signal Intens. x A CE-ESI-MS Intens. x B Time [min] 0.0 x C CE-ESI-MS with DEN-gas An overall lower background observed especially in the higher mass region Intens. x D Time [min] m/z 19 Kammeijer, G.S.M. et al. Manuscript in preparation
20 INCREASING SENSITIVITY OF GLYCAN ANALYSIS WITH CE-ESI-MS Improvement (total plasma N-glycome) 0 Kammeijer, G.S.M. et al. Manuscript in preparation
21 INCREASING SENSITIVITY OF GLYCAN ANALYSIS WITH CE-ESI-MS Improvement (total plasma N-glycome) 1 Kammeijer, G.S.M. et al. Manuscript in preparation
22 INCREASING SENSITIVITY OF GLYCAN ANALYSIS WITH CE-ESI-MS Improvement (total plasma N-glycome) 3x Kammeijer, G.S.M. et al. Manuscript in preparation
23 GLYCOMICS LOD DETERMINATION Sample PNGase F release Derivatization α,3 N-Acetylneuraminic acid α,6 N-Acetylneuraminic acid VS Purification and enrichment - Cotton HILIC Labeling - Addition of label with cationic charge CE-ESI-MS analysis 3 Haan, de N., et al., Anal. Chem., 015, 87 (16), pp , DOI: /acs.analchem.5b046cc
24 LOD EXPERIMENTAL SET-UP GLYCAN STANDARDS Dilution factor MALDI-TOF-MS Volume 1 µl Dilution series CE-ESI-MS Injection volume 44 nl Concentration Amount used Concentration (addition of LE, 9:1) Injected amount fmol/µl 1000 fmol 900 fmol/µl 39.6 fmol fmol/µl 100 fmol 90 fmol/µl 3.96 fmol 0 50 fmol/µl 50 fmol 45 fmol/µl 1.98 fmol fmol/µl 10 fmol 9 fmol/µl fmol 00 5 fmol/µl 5 fmol 4.5 fmol/µl fmol fmol/µl 1 fmol 0.9 fmol/µl fmol fmol/µl 0.5 fmol 0.45 fmol/µl 0.00 fmol fmol/µl 0.1 fmol 0.09 fmol/µl fmol 4 Kammeijer, G.S.M. et al. Manuscript in preparation
25 Relative peak area LOD EXPERIMENTAL SET-UP GLYCAN STANDARDS 000x CE-ESI-MS 1000x CE-ESI-MS 00x CE-ESI-MS 100x CE-ESI-MS 0x CE-ESI-MS 10x CE-ESI-MS 1x CE-ESI-MS 1x MALDI-TOF-MS 10x MALDI-TOF-MS 0x MALDI-TOF-MS 000x CE-ESI-MS 1000x CE-ESI-MS 00x CE-ESI-MS 100x CE-ESI-MS 0x CE-ESI-MS 10x CE-ESI-MS 1x CE-ESI-MS 1x MALDI-TOF-MS 10x MALDI-TOF-MS 0x MALDI-TOF-MS 70% 60% 50% 40% 30% 0% 10% 00% 5 Kammeijer, G.S.M. et al. Manuscript in preparation
26 LOD EXPERIMENTAL SET-UP GLYCAN STANDARDS MALDI-TOF-MS Dilution series Dilution series CE-ESI-MS MALDI-TOF-MS CE-ESI-MS Volume µl µl Injectionvolume volume nl nl Dilution Dilution factor Concentration Concentration Amount used used Concentration Concentration Injected Injected amount factor (addition of LE; 9:1) amount pmol/µl 1.8 pmol 3.83 pmol/µl 169 fmol fmol/µl 000 fmol 900 fmol/µl 39.6 fmol pmol/µl 100 fmol/µl 6.39 pmol 00 fmol 1.9 pmol/µl 90 fmol/µl 84.3 fmol 3.96 fmol pmol/µl 50 fmol/µl.56 pmol 100 fmol 0.77 pmol/µl 45 fmol/µl 33.7 fmol 1.98 fmol pmol/µl 10 fmol/µl 1.8 pmol 0 fmol 0.38 pmol/µl 9 fmol/µl 16.9 fmol fmol pmol/µl 0.64 pmol 0.19 pmol/µl 8.4 fmol 00 5 fmol/µl 10 fmol 4.5 fmol/µl fmol pmol/µl 0.6 pmol 0.07 pmol/µl 3. fmol fmol/µl fmol 0.9 fmol/µl fmol pmol/µl 0.5 fmol/µl 0.13 pmol 1 fmol 0.04 pmol/µl 0.45 fmol/µl 1.6 fmol 0.00 fmol fmol/µl 0. fmol 0.09 fmol/µl fmol CE-ESI-MS with DEN-gas shows a ~100x higher sensitivity compared to MALDI-TOF-MS 6 Kammeijer, G.S.M. et al. Manuscript in preparation
27 7 OUTLINE GLYCAN CHARACTERIZATION IMPROVING SENSITIVITY? IMPROVING SEPARATION 7 G.S.M. Kammeijer - g.s.m.kammeijer@lumc.nl
28 OPTIMIZATION DYNAMIC NEUTRALLY COATED CAPILLARY 8 BGE COMPOSITION 10 % AA 0 % AA 0 % AA 10 % MeOH TEMPERATURE COOLANT 15 C 0 C 5 C VOLTAGE 0 kv 5 kv 30 kv Flow 0 psi 0.3 psi 0.5 psi
29 TPNG PROFILE WITH CE-ESI-MS NEUTRALS Intens. x Dynamic Coated Neutral Capillary Injection: 5 psi 60sec (40 nl) Time [min] Dynamic neutrally coated BFS capillary, 90 cm 30µm id, BGE: 10 % AA, capillary coolant: 5 C, voltage: 0 kv, injection sample: 5 psi, 60 sec (40 nl) Intens. x Static Coated Neutral Capillary Injection: 1 psi 60sec (9 nl) Time [min] Static neutrally coated BFS capillary, 90 cm 30µm id, BGE: 10 % AA, capillary coolant: 5 C, voltage: 0 kv, injection sample: 1 psi, 60 sec (9 nl) 9 Kammeijer, G.S.M. et al. Manuscript in preparation
30 TPNG PROFILE WITH CE-ESI-MS NEUTRALS Intens. x Dynamic Coated Neutral Capillary Injection: 5 psi 60sec (40 nl) Time [min] Intens. x Static Coated Neutral Capillary Injection: 1 psi 60sec (9 nl) Time [min] 30 Kammeijer, G.S.M. et al. Manuscript in preparation
31 Intens. x x10 6 x10 6 x ZERO-FLOW PRINCIPLE 10 min 0 min 30 min 0 x min Time [min] 31 Kammeijer, G.S.M. et al. Manuscript in preparation
32 HIGH MANNOSE EFFECT OF ZERO-FLOW ON THE SEPARATION OF ISOMERIC N- GLYCANS GALACTOSYLATION Intensx min Intensx min Intensx min Intensx min Time [min] Intensx min Time [min] Intensx Time [min] 10 min 1.00 VS Time [min] Time [min] Time [min] Intensx min Intensx min VS Intensx min VS Time [min] Time [min] Time [min] Intensx min VS Intensx min Intensx min 0.00 Intensx min Time [min] 0.00 Intensx min Time [min] Intensx Time [min] 50 min Time [min] Time [min] Time [min]
33 EFFECT OF ZERO-FLOW ON THE SEPARATION OF ISOMERIC N- GLYCANS BISECTION/GALACTOSYLATION SIALYLATION Intensx10 4 Intensx10 4 Intensx10 4 Intensx10 4 Intensx min min min min min Time [min] Time [min] Time [min] Time [min] Time [min] VS VS Intensx Intensx Intensx Intensx Intensx min Time [min] 10 min Time [min] 0 min Time [min] 30 min Time [min] 50 min Time [min] Intensx Intensx Intensx Intensx Intensx min Time [min] 10 min Time [min] 0 min Time [min] 30 min Time [min] 50 min Time [min] VS
34 34 GLYCAN ANALYSIS WITH CESI-MS IN POSITIVE IONIZATION MODE o Glycan analysis is possible after derivatization and labeling o CE-ESI-MS found to be ~100 times more sensitive than MALDI-TOF-MS o CE-ESI-MS shows great potential for characterization of N-linked glycosylation in complex mixtures o Usage of static coated neutral capillary shows high potential o Isomeric separation visible for several N-glycan species Center for Proteomics and Metabolomics June 15, 016
35 35 PERSPECTIVES Characterization of various analytes o Investigation usage for biopharmaceutical characterization o Investigate the usage for limited sample amounts o Further explore the potential of isomeric separation using zero-flow o Comparison between positive and negative ionization mode o Investigate different labels o Identification of the different isomers
36 ACKNOWLEDGEMENTS Center for Proteomics and Metabolomics Manfred Wuhrer Noortje de Haan Sander Wagt Pablo Mohaupt Jan Nouta Karli Reiding Gerda Vreeker Bas Jansen Ludger Ltd. Rad Kozak Daniel Spencer Richard Gardner Darryl Fernandes Department of Chemistry & Chemical Biology Alexander Ivanov
Higher throughput glycosylation analysis of biopharmaceuticals by mass spectrometry
Higher throughput glycosylation analysis of biopharmaceuticals by mass spectrometry Manfred Wuhrer Center for Proteomics and Metabolomics CASSS AT EUROPE, BARCELONA, MARCH 8, 18 GLYCOSYLATION Highly prevalent
More informationSupporting Information for MassyTools-assisted data analysis of total serum N-glycome changes associated with pregnancy
Supporting Information for MassyTools-assisted data analysis of total serum N-glycome changes associated with pregnancy Bas C. Jansen 1, Albert Bondt 1,2, Karli R. Reiding 1, Coen J. de Jong 1, David Falck
More informationApplying a Novel Glycan Tagging Reagent, RapiFluor-MS, and an Integrated UPLC-FLR/QTof MS System for Low Abundant N-Glycan Analysis
Applying a Novel Glycan Tagging Reagent, RapiFluor-MS, and an Integrated UPLC-FLR/QTof MS System for Low Abundant N-Glycan Analysis Ying Qing Yu Waters Corporation, Milford, MA, USA APPLICATION BENEFITS
More informationApplication Note. Abstract. Author. Biotherapeutics & Biosimilars. Sonja Schneider Agilent Technologies, Inc. Waldbronn, Germany
Sensitive and Reproducible Glycan Analysis of Human Immunoglobulin G The Agilent 1260 Infi nity Bio-inert Quaternary LC System with an Agilent AdvanceBio 2.7 µm Glycan Mapping Column and Fluorescence Detection
More informationSupporting information
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2014 Supporting information Glycan Reductive Isotope-coded Amino Acid Labeling (GRIAL) for Mass Spectrometry-based
More informationGlycanPac AXR-1 Columns
CHRMATGRAPHY GlycanPac AXR- Columns For High Resolution Glycan Analysis Product Specifications The Thermo Scientific GlycanPac AXR- columns are highperformance, silica-based HPLC columns for simultaneous
More informationMass Spectrometry at the Laboratory of Food Chemistry. Edwin Bakx Laboratory of Food Chemistry Wageningen University
Mass Spectrometry at the Wageningen University Mass Spectrometry at the 3 UPLC/CE - ESI - Ion trap MS systems UPLC Thermo Acella with a Velos or VelosPro CE Beckman PA800 with a Thermo VelosPro 1 UPLC-
More informationSeparation of 15 Underivatized Saccharide and Sialic Acid USP Standards
Application Note Raw Material & Media Analysis Separation of Underivatized Saccharide and Sialic Acid USP Standards Using an Agilent AdvanceBio MS Spent Media Column with TOF MS Detection Author Richard
More informationBarry Boyes 1,2, Shujuan Tao 2, and Ron Orlando 2
Barry Boyes 1,2, Shujuan Tao 2, and Ron Orlando 2 1 Advanced Materials Technology, Inc. Wilmington, DE USA 2 Complex Carbohydrate Research Center University of Georgia, Athens, GA USA bboyes@advanced-materials-tech.com
More informationCurrent Glycoprotein Analysis. Glycan Characterization: Oligosaccharides. Glycan Analysis: Sample Preparation. Glycan Analysis: Chromatography
Bio Day DENMARK MARCH 2013 Analysis of N-linked Glycans of GlycoProteins marleen_van_wingerden@waters.com Agenda Importance of Glycan Analysis Current Glycoprotein Analysis Glycan Characterization: Oligosaccharides
More informationSeparation of 2AA-Labeled N-Linked Glycans from Glycoproteins on a High Resolution Mixed-Mode Column
Separation of 2AA-Labeled N-Linked Glycans from Glycoproteins on a High Resolution Mixed-Mode Column Udayanath Aich, Julian Saba, Xiaodong Liu, Jeff Rohrer, Jim Thayer and Chris Pohl, Thermo Fisher Scientific,
More informationThermo Fisher Scientific, Sunnyvale, CA, USA; 2 Thermo Fisher Scientific, San Jose, CA, USA
An Ultra High Resolution Glycan Column for Isomeric Separation and the Structural Identification of Labeled N-Glycans from Proteins Including Antibodies Udayanath Aich, 1 Julian Saba, 2 Rosa Viner, 2 Shanhua
More informationNew Instruments and Services
New Instruments and Services http://planetorbitrap.com/orbitrap fusion Combining the best of quadrupole, Orbitrap, and ion trap mass analysis in a revolutionary Tribrid architecture, the Orbitrap Fusion
More informationA Novel HILIC Column for High Speed N-linked Glycan Analysis
A Novel HILIC for High Speed N-linked Glycan Analysis Application Note Biotherapeutics and Biosimilars Authors James Martosella, Oscar Potter, Danny Mancheno, and Jia Liu Agilent Technologies, Inc. Introduction
More informationComparison of mass spectrometers performances
Comparison of mass spectrometers performances Instrument Mass Mass Sensitivity resolution accuracy Quadrupole 1 x 10 3 0.1 Da* 0.5-1.0 pmol DE-MALDI 2 x 10 4 20 ppm 1-10 fmol peptide 1-5 pmol protein Ion
More informationAnalysis of N-Linked Glycans from Coagulation Factor IX, Recombinant and Plasma Derived, Using HILIC UPLC/FLR/QTof MS
Analysis of N-Linked Glycans from Coagulation Factor IX, Recombinant and Plasma Derived, Using HILIC UPLC/FLR/QTof MS Ying Qing Yu Waters Corporation, Milford, MA, U.S. A P P L I C AT ION B E N E F I T
More informationMass Spectrometry. Actual Instrumentation
Mass Spectrometry Actual Instrumentation August 2017 See also http://www.uni-bielefeld.de/chemie/analytik/ms f additional infmation 1. MALDI TOF MASS SPECTROMETRY ON THE ULTRAFLEX 2 2. ESI MASS SPECTROMETRY
More informationA computational framework for discovery of glycoproteomic biomarkers
A computational framework for discovery of glycoproteomic biomarkers Haixu Tang, Anoop Mayampurath, Chuan-Yih Yu Indiana University, Bloomington Yehia Mechref, Erwang Song Texas Tech University 1 Goal:
More informationProfiling the Distribution of N-Glycosylation in Therapeutic Antibodies using the QTRAP 6500 System
Profiling the Distribution of N-Glycosylation in Therapeutic Antibodies using the QTRAP 6500 System Scheduled MRM Pro Algorithm for Increased Efficiency of Targeted Detection Jenny Albanese 1, Christie
More informationThank you for joining us! Our session will begin shortly Waters Corporation 1
UPLC and HPLC Separation Strategies for Successful Characterization of Glycans Derived from Therapeutic Proteins Thank you for joining us! Our session will begin shortly 2013 Waters Corporation 1 Friendly
More informationTECHNICAL BULLETIN. R 2 GlcNAcβ1 4GlcNAcβ1 Asn
GlycoProfile II Enzymatic In-Solution N-Deglycosylation Kit Product Code PP0201 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description Glycosylation is one of the most common posttranslational
More informationAnalysis of Glycopeptides Using Porous Graphite Chromatography and LTQ Orbitrap XL ETD Hybrid MS
Analysis of Glycopeptides Using Porous Graphite Chromatography and LTQ Orbitrap XL ETD Hybrid MS Terry Zhang, Rosa Viner, Zhiqi Hao, Vlad Zabrouskov, Thermo Fisher Scientific, San Jose, CA, USA Introduction
More informationAnalysis of Choline and its Metabolites on the Finnigan LTQ
Analysis of Choline and its Metabolites on the Finnigan LTQ Gargi Choudhary Diane Cho Thermo Electron, San Jose, CA Abstracted from a poster presented at Montreux 3 LC/MS Symposium, with co-authors: Brad
More informationStructural Analysis of Labeled N-Glycans from Proteins by LC-MS/MS Separated Using a Novel Mixed-Mode Stationary Phase
Structural Analysis of Labeled N-Glycans from Proteins by LC-MS/MS Separated Using a Novel Mixed-Mode Stationary Phase Udayanath Aich 1, Julian Saba 2, Xiaodong Liu 1, Srinivasa Rao 1, and Chris Pohl 1
More informationHigh Throughput Accurate Mass Screening of Monoclonal Antibodies
High Throughput Accurate Mass Screening of Monoclonal Antibodies Jim Blasberg April 28, 211 sigma-aldrich.com Life Science and High Technology Center Life Science and High Technology Center2 Sigma-Aldrich,
More informationAdvances in Hybrid Mass Spectrometry
The world leader in serving science Advances in Hybrid Mass Spectrometry ESAC 2008 Claire Dauly Field Marketing Specialist, Proteomics New hybrids instruments LTQ Orbitrap XL with ETD MALDI LTQ Orbitrap
More informationDetermination of Benzodiazepines in Urine by CE-MS/MS
Determination of Benzodiazepines in Urine by CE-MS/MS Application ote Forensic Toxicology Authors audimir Lucio do Lago Department of Fundamental Chemistry, Institute of Chemistry University of São Paulo,
More informationProfiling of Endogenous Metabolites Using Time-of-Flight LC/MS with Ion Pair Reverse Phase Chromatography
Profiling of Endogenous Metabolites Using Time-of-Flight LC/MS with Ion Pair Reverse Phase Chromatography Application Note Metabolomics Author Yuqin Dai Agilent Technologies, Inc. 31 Stevens Creek Blvd,
More informationNew Instruments and Services
New Instruments and Services Liwen Zhang Mass Spectrometry and Proteomics Facility The Ohio State University Summer Workshop 2016 Thermo Orbitrap Fusion http://planetorbitrap.com/orbitrap fusion Thermo
More informationOnline 2D-LC Analysis of Complex N-Glycans in Biopharmaceuticals Using the Agilent 1290 Infinity 2D-LC Solution
Online D-LC Analysis of Complex N-Glycans in Biopharmaceuticals Using the Agilent 19 Infinity D-LC Solution Comprehensive and Multiple Heart-Cutting D-LC Analysis for Highest Resolution Application Note
More informationDetermination of Amphetamine and Derivatives in Urine
Determination of Amphetamine and Derivatives in Urine Using a Modified QuEChERS and Capillary Electrophoresis Tandem Mass Spectrometry Analysis Application Note Authors Vagner B. dos Santos and Claudimir
More informationGlycan and Monosaccharide Workshop Eoin Cosgrave David Wayland Bill Warren
Glycan and Monosaccharide Workshop Eoin Cosgrave David Wayland Bill Warren 2012 Waters Corporation 1 Requests and Questions Optimised sample prep protocol to reduce sample preparation time How can I detect
More informationHigh Resolution Glycopeptide Mapping of EPO Using an Agilent AdvanceBio Peptide Mapping Column
High Resolution Glycopeptide Mapping of EPO Using an Agilent AdvanceBio Peptide Mapping Column Application Note BioPharma Authors James Martosella, Phu Duong, and Alex Zhu Agilent Technologies, Inc. Abstract
More informationBioanalytical Quantitation of Biotherapeutics Using Intact Protein vs. Proteolytic Peptides by LC-HR/AM on a Q Exactive MS
Bioanalytical Quantitation of Biotherapeutics Using Intact Protein vs. Proteolytic Peptides by LC-HR/AM on a Q Exactive MS Jenny Chen, Hongxia Wang, Zhiqi Hao, Patrick Bennett, and Greg Kilby Thermo Fisher
More informationMALDI Imaging Drug Imaging Detlev Suckau Head of R&D MALDI Bruker Daltonik GmbH. December 19,
MALDI Imaging Drug Imaging Detlev Suckau Head of R&D MALDI Bruker Daltonik GmbH December 19, 2014 1 The principle of MALDI imaging Spatially resolved mass spectra are recorded Each mass signal represents
More informationMS/MS as an LC Detector for the Screening of Drugs and Their Metabolites in Race Horse Urine
Application Note: 346 MS/MS as an LC Detector for the Screening of Drugs and Their Metabolites in Race Horse Urine Gargi Choudhary and Diane Cho, Thermo Fisher Scientific, San Jose, CA Wayne Skinner and
More informationLudger Guide to Sialylation: II. Highly Sialylated Glycoproteins
Ludger Guide to Sialylation: II Highly Sialylated Glycoproteins Ludger has over 15 years experience providing products and services for the biopharmaceutical industry and in that time we have noticed that
More informationAutomating Mass Spectrometry-Based Quantitative Glycomics using Tandem Mass Tag (TMT) Reagents with SimGlycan
PREMIER Biosoft Automating Mass Spectrometry-Based Quantitative Glycomics using Tandem Mass Tag (TMT) Reagents with SimGlycan Ne uaca2-3galb1-4glc NAcb1 6 Gal NAca -Thr 3 Ne uaca2-3galb1 Ningombam Sanjib
More informationSimGlycan. A high-throughput glycan and glycopeptide data analysis tool for LC-, MALDI-, ESI- Mass Spectrometry workflows.
PREMIER Biosoft SimGlycan A high-throughput glycan and glycopeptide data analysis tool for LC-, MALDI-, ESI- Mass Spectrometry workflows SimGlycan software processes and interprets the MS/MS and higher
More informationAn Alternative Approach: Top-Down Bioanalysis of Intact Large Molecules Can this be part of the future? Lecture 8, Page 27
An Alternative Approach: Top-Down Bioanalysis of Intact Large Molecules Can this be part of the future? Lecture 8, Page 27 Top-down HRAM Bioanalysis of Native Proteins/Molecules Relative Abundance 100
More informationJohn Haselden 1, Gordon Dear 1, Jennifer H. Granger 2, and Robert S. Plumb 2. 1GlaxoSmithKline, Ware, UK; 2 Waters Corporation, Milford, MA, USA
Challenges of Polar Compound Analysis Routine LC/MS-based analyses usually rely on using reversed-phase column chemistries (e.g. C 18, C 8, C 4 ) in their LC methodology. However, one of the shortcomings
More informationAnalysis of 2-Aminobenzamide (AB) Labeled Glycans Using HPLC with Fluorescence Detection
Technical Note 19 Analysis of 2-Aminobenzamide (AB) Labeled Glycans Using HPLC with Fluorescence Detection Introduction The study of a protein s glycosylation can reveal highly valuable diagnostic information.
More informationThe Road to Glycan Analysis Without Compromise WCBP 2015 Waters Technical Seminar Jan 27, 2015 Washington, DC
The Road to Glycan Analysis Without Compromise WCBP 2015 Waters Technical Seminar Jan 27, 2015 Washington, DC 2015 Waters Corporation 1 Today s Agenda Overview and Introduction RapiFluor-MS N-Glycan Labeling:
More informationFOURIER TRANSFORM MASS SPECTROMETRY
FOURIER TRANSFORM MASS SPECTROMETRY https://goo.gl/vx3ogw FT-ICR Theory Ion Cyclotron Motion Inward directed Lorentz force causes ions to move in circular orbits about the magnetic field axis Alan G. Marshall,
More informationCan we learn something new about peptide separations after 40 years of RP and HILIC chromatography? Martin Gilar April 12, MASSEP 2016
Can we learn something new about peptide separations after 40 years of RP and HILIC chromatography? Martin Gilar April 12, MASSEP 2016 2015 Waters Corporation 1 Overview 1. 2D RP RP LC of peptides 2. RP-LC
More informationLOCALISATION, IDENTIFICATION AND SEPARATION OF MOLECULES. Gilles Frache Materials Characterization Day October 14 th 2016
LOCALISATION, IDENTIFICATION AND SEPARATION OF MOLECULES Gilles Frache Materials Characterization Day October 14 th 2016 1 MOLECULAR ANALYSES Which focus? LOCALIZATION of molecules by Mass Spectrometry
More informationThermo Scientific. GlycanPac AXR-1. Column Product Manual. P/N: April, Part of Thermo Fisher Scientific
Thermo Scientific GlycanPac AXR-1 Column Product Manual P/N: April, 2014 Part of Thermo Fisher Scientific Product Manual for GlycanPac AXR-1 Columns GlycanPac AXR-1, 1.9 µm, Analytical, 2.1 x 150 mm (P/N
More informationDetermination of B-vitamins in Energy Drinks by CE/MS/MS
Application ote Food Determination of B-vitamins in Energy Drinks by E/MS/MS Authors laudimir Lucio do Lago and Zuzana ieslarová Department of Fundamental hemistry, Institute of hemistry, University of
More informationGlycosylation analysis of blood plasma proteins
Glycosylation analysis of blood plasma proteins Thesis booklet Eszter Tóth Doctoral School of Pharmaceutical Sciences Semmelweis University Supervisor: Károly Vékey DSc Official reviewers: Borbála Dalmadiné
More informationMASS SPECTROMETRY BASED METABOLOMICS. Pavel Aronov. ABRF2010 Metabolomics Research Group March 21, 2010
MASS SPECTROMETRY BASED METABOLOMICS Pavel Aronov ABRF2010 Metabolomics Research Group March 21, 2010 Types of Experiments in Metabolomics targeted non targeted Number of analyzed metabolites is limited
More informationFlow-Through Electron Capture Dissociation in a novel Branched RF Ion Trap
Flow-Through Electron Capture Dissociation in a novel Branched RF Ion Trap Takashi Baba, J. Larry Campbell, Yves Le Blanc, Jim. W. Hager and Bruce A. Thomson ASMS, June 18 / 2014 1 2014 AB SCIEX Trapping
More informationChallenges in Separation Technologies
Challenges in Separation Technologies Erin Shammel Baker Xing Zhang, Yehia Ibrahim, Matthew Monroe, Dennis Mehinagic, Justin Teeguarden, Thomas Metz and Richard D. Smith Pacific Northwest National Laboratory
More informationBiotherapeutics. Biopharmaceutical Sciences Group Waters Corporation Waters Corporation 1
Characterization of the Higher-Order Structure of Biotherapeutics Biopharmaceutical Sciences Group Waters Corporation 2011 Waters Corporation 1 FDA desire for specific innovations in biotherapeutic analysis
More informationQSTAR Operation. May 3, Bob Seward
QSTAR Operation May 3, 2005 Bob Seward Presentation CD Contents QSTAR Hardware QSTAR Schematic 10-7 torr QSTAR Schematic Chernushevich et al, JMS 2001, 36: 849-865 Curtain Plate ESI Source ESI Source Curtain
More informationOligosaccharide Analysis by High-Performance Anion- Exchange Chromatography with Pulsed Amperometric Detection
Oligosaccharide Analysis by High-Performance Anion- Exchange Chromatography with Pulsed Amperometric Detection Jeff Rohrer, Ph.D. Director, Applications Development, Dionex Products 1 The world leader
More informationLC/MS Method for Comprehensive Analysis of Plasma Lipids
Application Note omics LC/MS Method for Comprehensive Analysis of Plasma s Authors Tomas Cajka and Oliver Fiehn West Coast Metabolomics Center, University of California Davis, 451 Health Sciences Drive,
More informationGlycosylation analyses of recombinant proteins by LC-ESI mass spectrometry
Glycosylation analyses of recombinant proteins by LC-ESI mass spectrometry Dr Malin Bäckström Mammalian Protein Expression Core Facility P4EU meeting Porto Nov 11-12, 2013 MPE - A tissue culture facility
More informationfor the Identification of Phosphorylated Peptides
Application of a Data Dependent Neutral-Loss Experiment on the Finnigan LTQ for the Identification of Phosphorylated Peptides Gargi Choudhary Diane Cho Thermo Electron, San Jose, CA Abstracted from posters
More informationAnalyze Biogenic Amines in Salmon and Shrimp by Capillary Electrophoresis-Tandem MS
Analyze Biogenic Amines in Salmon and Shrimp by Capillary Electrophoresis-Tandem MS Application ote Food Testing and Agriculture Authors Claudimir Lucio do Lago, Vagner B. dos Santos, and Denis T. R. Vidal
More informationPHOSPHOPEPTIDE ANALYSIS USING IMAC SAMPLE PREPARATION FOLLOWED BY MALDI-MS and MALDI PSD MX
PHOSPHOPEPTIDE ANALYSIS USING IMAC SAMPLE PREPARATION FOLLOWED BY MALDI-MS and MALDI PSD MX E. Claude 1, E. Emekwue 2, M. Snel 1, T. McKenna 1 and J. Langridge 1 1: Waters Corporation, Manchester, UK 2:
More informationApplication Note # ET-17 / MT-99 Characterization of the N-glycosylation Pattern of Antibodies by ESI - and MALDI mass spectrometry
Bruker Daltonics Application Note # ET-17 / MT-99 Characterization of the N-glycosylation Pattern of Antibodies by ESI - and MALDI mass spectrometry Abstract Analysis of the N-glycosylation pattern on
More informationComparison of Relative Quantification of Monoclonal Antibody N-glycans Using Fluorescence and MS Detection
Comparison of Relative Quantification of Monoclonal ntibody N-glycans Using Fluorescence and MS Detection pplication Note iotherapeutics & iologics uthors scar Potter and Gregory Staples gilent Technologies,
More informationSYNAPT G2-S High Definition MS (HDMS) System
SYNAPT G2-S High Definition MS (HDMS) System High performance, versatility, and workflow efficiency of your MS system all play a crucial role in your ability to successfully reach your scientific and business
More informationN-Glycan Sequencing Kit
PROTEIN TOOLS N-Glycan Sequencing Kit Instruction Manual NEB #E577S 2 reactions Version 1. 1/18 be INSPIRED drive DISCOVERY stay GENUINE This product is intended for research purposes only. This product
More informationPhospholipid characterization by a TQ-MS data based identification scheme
P-CN1716E Phospholipid characterization by a TQ-MS data based identification scheme ASMS 2017 MP-406 Tsuyoshi Nakanishi 1, Masaki Yamada 1, Ningombam Sanjib Meitei 2, 3 1 Shimadzu Corporation, Kyoto, Japan,
More informationamazon speed Innovation with Integrity Ion Trap Performance Beyond Imagination Ion Trap MS
amazon speed Ion Trap Performance Beyond Imagination Innovation with Integrity Ion Trap MS amazon Turning Speed into Solutions The amazon speed ion trap series sets new analytical standards for proteomics
More informationIsomeric Separation of Permethylated Glycans by Porous Graphitic Carbon (PGC)-LC-MS/MS at High- Temperatures
Supplementary Information Isomeric Separation of Permethylated Glycans by Porous Graphitic Carbon (PGC)-LC-MS/MS at High- Temperatures Shiyue Zhou 1, Yifan Huang 1, Xue Dong 1, Wenjing Peng 1, Lucas Veillon
More information[ APPLICATION NOTE ] High Sensitivity Intact Monoclonal Antibody (mab) HRMS Quantification APPLICATION BENEFITS INTRODUCTION WATERS SOLUTIONS KEYWORDS
Yun Wang Alelyunas, Henry Shion, Mark Wrona Waters Corporation, Milford, MA, USA APPLICATION BENEFITS mab LC-MS method which enables users to achieve highly sensitive bioanalysis of intact trastuzumab
More informationCESI-MS of Intact Proteins
CESI-MS of Intact Proteins Rob Haselberg, Chitra K. Ratnayake, Gerhardus J. de Jong, Govert W. Somsen. iomolecular nalysis, Utrecht University, Universiteitsweg 99, 58 CG, The Netherlands. eckman Coulter,
More informationSupporting information
Supporting information Figure legends Supplementary Table 1. Specific product ions obtained from fragmentation of lithium adducts in the positive ion mode comparing the different positional isomers of
More informationMetabolomics: quantifying the phenotype
Metabolomics: quantifying the phenotype Metabolomics Promises Quantitative Phenotyping What can happen GENOME What appears to be happening Bioinformatics TRANSCRIPTOME What makes it happen PROTEOME Systems
More informationAnalyzing Phenyl Ureas and Carbamates with HPLC/API-MS. Presented by Chen-Kai Meng, Ph.D. Applications Chemist
Analyzing Phenyl Ureas and Carbamates with HPLC/API-MS Presented by Chen-Kai Meng, Ph.D. Applications Chemist Outline Overview of ESI, APPI, and APCI sources Comparing ESI, APPI, and APCI results Identifying
More informationLC/MS Analysis of Various Hydrophilic Compounds Using a Polymer-Based Amino Column - Shodex TM HILICpak TM VG-50 2D
LC/MS Analysis of Various Hydrophilic Compounds Using a Polymer-Based Amino Column - Shodex TM HILICpak TM VG-50 2D Introduction Components of pharmaceutical products and food products often include high
More informationIntroduction to Proteomics 1.0
Introduction to Proteomics 1.0 CMSP Workshop Pratik Jagtap Managing Director, CMSP Objectives Why are we here? For participants: Learn basics of MS-based proteomics Learn what s necessary for success using
More informationBlotGlyco Glycan purification and labeling kit
Aug. 14th, 2012 BlotGlyco Glycan purification and labeling kit S-BIO Business Division SUMITOMO BAKELITE CO., LTD. 1 BlotGlyco kit Biological sample Glycan Hz BlotGlyco :Label MS HPLC 2 BlotGlyco kit is
More informationMass spectra of peptides and proteins - and LC analysis of proteomes Stephen Barnes, PhD
Mass spectra of peptides and proteins - and LC analysis of proteomes Stephen Barnes, PhD 4-7117 sbarnes@uab.edu Overview A mass spectrum Electrospray MS Analysis of intact proteins Molecular weight calculations
More informationSample Preparation is Key
PLOS ONE DOI: 10.1371/journal.pone.0117232 February 6, 2015 Presented by Katie Gibbs Sample Preparation is Key Sample extraction and instrumental analysis methods are well documented in metabolomics. Understanding
More informationMetabolite identification in metabolomics: Metlin Database and interpretation of MSMS spectra
Metabolite identification in metabolomics: Metlin Database and interpretation of MSMS spectra Jeevan K. Prasain, PhD Department of Pharmacology and Toxicology, UAB jprasain@uab.edu Outline Introduction
More informationAutomated Sample Preparation/Concentration of Biological Samples Prior to Analysis via MALDI-TOF Mass Spectroscopy Application Note 222
Automated Sample Preparation/Concentration of Biological Samples Prior to Analysis via MALDI-TOF Mass Spectroscopy Application Note 222 Joan Stevens, Ph.D.; Luke Roenneburg; Tim Hegeman; Kevin Fawcett
More informationA NOVEL METHOD OF M/Z DRIFT CORRECTION FOR OA-TOF MASS SPECTROMETERS BASED ON CONSTRUCTION OF LIBRARIES OF MATRIX COMPONENTS.
A NOVEL METHOD OF M/Z DRIFT CORRECTION FOR OA-TOF MASS SPECTROMETERS BASED ON CONSTRUCTION OF LIBRARIES OF MATRIX COMPONENTS. Martin R Green*, Keith Richardson, John Chipperfield, Nick Tomczyk, Martin
More informationExtended Mass Range Triple Quadrupole for Routine Analysis of High Mass-to-charge Peptide Ions
Extended Mass Range Triple Quadrupole for Routine Analysis of High Mass-to-charge Peptide Ions Application Note Targeted Proteomics Authors Linfeng Wu, Christine A. Miller, Jordy Hsiao, Te-wei Chu, Behrooz
More informationAutomated Detection and Identification of N- and O-glycopeptides
Discovering the Subtleties of Sugars June 10 th 14 th, 2013, Potsdam, Germany 85 Automated Detection and Identification of N- and O-glycopeptides Peter Hufnagel *, Anja Resemann, Wolfgang Jabs, Kristina
More informationMore structural information with MS n
PRODUCT SPECIFICATIONS The LTQ XL linear ion trap mass spectrometer More structural information with MS n The LTQ XL linear ion trap mass spectrometer delivers more structural information faster and with
More informationA CZE Method for the Identification of Therapeutic Antibodies and Quality Control of Infusion Bags at the Hospital
CE in the biotechnology and pharmaceutical industries, 17th Symposium on the Practical Applications for the Analysis of Proteins, Nucleotides and Small Molecules, September 20, 2015 - Thursday, September
More informationA systematic investigation of CID Q-TOF-MS/MS collision energies to allow N- and O-glycopeptide identification by LC-MS/MS
A systematic investigation of CID Q-TO-MS/MS collision energies A systematic investigation of CID Q-TO-MS/MS collision energies to allow N- and O-glycopeptide identification by LC-MS/MS Abstract The MS
More informationA Fully Integrated Workflow for LC-MS/MS Analysis of Labeled and Native N-Linked Glycans Released From Proteins
A Fully Integrated Workflow for LC-MS/MS Analysis of Labeled and Native N-Linked Glycans Released From Proteins Udayanath Aich, 1 Julian Saba, 2 Xiaodong Liu, 1 Srinivasa Rao, 1 Yury Agroskin, 1 and Chris
More informationREDOX PROTEOMICS. Roman Zubarev.
REDOX PROTEOMICS Roman Zubarev Roman.Zubarev@ki.se Physiological Chemistry I, Department for Medical Biochemistry & Biophysics, Karolinska Institutet, Stockholm What is (RedOx) Proteomics? Proteomics -
More informationO O H. Robert S. Plumb and Paul D. Rainville Waters Corporation, Milford, MA, U.S. INTRODUCTION EXPERIMENTAL. LC /MS conditions
Simplifying Qual/Quan Analysis in Discovery DMPK using UPLC and Xevo TQ MS Robert S. Plumb and Paul D. Rainville Waters Corporation, Milford, MA, U.S. INTRODUCTION The determination of the drug metabolism
More informationApplication Note # LCMS-89 High quantification efficiency in plasma targeted proteomics with a full-capability discovery Q-TOF platform
Application Note # LCMS-89 High quantification efficiency in plasma targeted proteomics with a full-capability discovery Q-TOF platform Abstract Targeted proteomics for biomarker verification/validation
More informationIon Exchange and Reversed Phase interactions in selective Bio-SPME extractions of designer drugs
Ion Exchange and Reversed Phase interactions in selective Bio-SPME extractions of designer drugs Frank Michel, Craig R. Aurand, Robert E. Shirey, Yong Chen, Leonard M. Sidisky sigma-aldrich.com/analytical
More informationStructural Elucidation of N-glycans Originating From Ovarian Cancer Cells Using High-Vacuum MALDI Mass Spectrometry
PO-CON1347E Structural Elucidation of N-glycans Originating From Ovarian Cancer Cells Using High-Vacuum MALDI Mass Spectrometry ASMS 2013 TP-708 Matthew S. F. Choo 1,3 ; Roberto Castangia 2 ; Matthew E.
More informationElectron transfer dissociation (ETD)
Electron transfer dissociation (ETD) Electron transfer dissociation (ETD) Originally developed for FT-ICR MS as ECD ( C for capture) Now often used on other trapping instruments, even Q-TOFs Trick: Bringing
More informationMass Spectrometry Infrastructure
Mass Spectrometry Infrastructure Todd Williams, Ph.D. Director KU Mass Spectrometry and Analytical Proteomics Laboratory Mass Spectrometry Lab B025 Malott Hall Mission The Mass Spectrometry and analytical
More informationBreaking the Sensitivity Barrier in Triple Quad LC/MS Performance
Breaking the Sensitivity Barrier in Triple Quad LC/MS Performance Tim Schlabach Alex Mordehai Michael Zumwalt Page 1 Sensitivity enhancements for ESI-MS Reduce the column diameter! 30 fold gains in sensitivity
More informationQuantitative Analysis of Vit D Metabolites in Human Plasma using Exactive System
Quantitative Analysis of Vit D Metabolites in Human Plasma using Exactive System Marta Kozak Clinical Research Applications Group Thermo Fisher Scientific San Jose CA Clinical Research use only, Not for
More informationNovel Glycan Column Technology for the LC-MS Analysis of Labeled and Native N-Glycans Released from Proteins and Antibodies
Novel Glycan Column Technology for the LC-MS Analysis of Labeled and Native N-Glycans Released from Proteins and Antibodies Udayanath Aich, 1 Ilze Birznieks, 1 Julian Saba, 2 Xiaodong Liu, 1 Rosa Viner,
More informationIonization Methods. Neutral species Charged species. Removal/addition of electron(s) Removal/addition of proton(s)
Ionization Methods Neutral species Charged species Removal/addition of electron(s) M + e - (M +. )* + 2e - electron ionization Removal/addition of proton(s) M + (Matrix)-H MH + + (Matrix) - chemical ionization
More informationPHOTOCATALYTIC DECONTAMINATION OF CHLORANTRANILIPROLE RESIDUES IN WATER USING ZnO NANOPARTICLES. DR. A. RAMESH, Ph.D, D.Sc.,
PHOTOCATALYTIC DECONTAMINATION OF CHLORANTRANILIPROLE RESIDUES IN WATER USING ZnO NANOPARTICLES DR. A. RAMESH, Ph.D, D.Sc., raamesh_a@yahoo.co.in 1 OBJECTIVES Determination of persistence and photolysis
More informationMultiple Reaction Monitoring for Direct Quantitation of Intact Proteins using a Triple Quadrupole Mass Spectrometer
ELECTRONIC SUPPLEMENTARY INFORMATION Multiple Reaction Monitoring for Direct Quantitation of Intact Proteins using a Triple Quadrupole Mass Spectrometer Evelyn H. Wang 1, Peter C. Combe 2, and Kevin A.
More informationDetermination of Chlorophenoxyacetic Acid and Other Acidic Herbicides Using a QuEChERS Sample Preparation Approach and LC-MS/MS Analysis
Determination of Chlorophenoxyacetic Acid and Other Acidic Herbicides Using a QuEChERS Sample Preparation Approach and LC-MS/MS Analysis UCT Product Number: ECQUEU75CT-MP - Mylar pouch containing extraction
More information