New Instruments and Services

Transcription

1 New Instruments and Services

2 fusion

3 Combining the best of quadrupole, Orbitrap, and ion trap mass analysis in a revolutionary Tribrid architecture, the Orbitrap Fusion Tribrid mass spectrometer features: Resolution up to 450,000 to remove spectral interferences CID/HCD/ETD detected by the Ion Trap or Orbitrap at any level of MSn for maximum experimental flexibility Dynamic Scan Management architecture lets every user easily get the maximum information from every sample Quick polarity switch between positive and negative mode.

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5 The ParaCell is the new core technology for solarix XR, providing uncommon broadband ion stability resulting in resolution far superior to other detection schemes. Ultimate mass resolving power in excess of RP = 10,000,000 FTICR technology providing unrivaled mass accuracy Apollo II Ion Funnel Electrospray Source enabling high ion transmission efficiency over a broad mass range Hybrid Qh Front End supporting CASI for enrichment of low abundant species Optimized ion optics for sensitivity and mass range coverage Proprietary ParaCell ICR cell with patented SIDEKICK ion accumulation system for high detection efficiency. Nanobay E Acquisition electronics enabling data streaming for increased broad band resolution Wide range of fragmentation techniques including CID and ECD Dual ionization source: MALDI source with Smartbeam II laser and ESI source

6 Application of Orbitrap Fusion PROTEINS Bottom-up Proteomics Targeted Quantitation Top-Down Proteomics Intact Protein Characterization Post Translational Modification Analysis: Glycosylation Phosphorylation Application of 15T FT ICR High End Proteomics Studies (Top-down and Bottom-up workflows) PTM Investigation Accurate de novo sequencing Molecular Imaging of Tissue Distribution of Drugs, Metabolites, and Biomarkers Petroleum Product Analysis Complex Environmental Sample Analysis Metabolomics Research Relative Quantitation: SILAC TMT Label-Free GLYCANS Glycan Identification & Characterization 2-AA Quantitation Intact Glycoprotein Profiling PTM Glycosylation

7 New Services: 2D LC/MSMS for Protein Identification and PTM Study Targeted Proteomics Quantitation Top-down Proteomics MALDI Tissue Imaging

8 High ph Fractionation with Low ph Separation Waters Mclass 2D System Dionex D System $60/fraction on the orbitrap XL $75/fraction on the orbitrap Fusion Slide courtesy of Nilini S. Ranbaduge

9 Multi-dimensional Separation ph 10.0 ph 2.6 F1 $60/fraction on the orbitrap XL $75/fraction on the orbitrap Fusion Time (min) RT: F2 Relative Abundance Time (min) Fractionation (F) Time (min) Gradient separation Time (min) F15 9 Slide courtesy of Nilini S. Ranbaduge

10 RT: Targeted Proteomics Quantitation Relative Abundance Full Scan (SLVLfGlyTPSR )/ (SLVLC (CAM) TPSR) NL: 2.47E8 Base Peak m/z= MS 17625_1_rer un NL: 8.17E8 Base Peak MS 17625_1_Ta rget_ Time (min) MS/MS of (SLVLC (CAM) TPSR) 17625_1_Target_2 # RT: AV: 264 NL: 3.92E4 T: ITMS + c ESI d Full ms @cid35.00 [ ] MS/MS of (SLVLfGlyTPSR ) 17625_1_Target_2 # RT: AV: 40 NL: 1.03E3 T: ITMS + c ESI d Full ms @cid35.00 [ ] Relative Abundance Cys(CAM) Leu Val Thr Val Leu Cys(CAM) Thr Pro Ser m/z Relative Abundance Thr fgly Leu Val Val Leu fgly Thr m/z

11 Top down v.s. Bottom up Proteomics Circulation: Cardiovascular Genetics December 2011 vol. 4 no Trends in Biochemical Science; Volume 35, Issue 9, September 2010, Pages

12 Protein Identification by Top Down MS/MS Parents ions ( , 15+) Ac + 2Me + 2Ac + 2Me Histone H4 +1 Ac + 4Ac + 5Ac m/z ECD MS/MS c13+42 z64 c z50 z42 c c15+42 C24+70 c c c7+42 c c59+70 z20 c c c c26+70 c12+42 c52+70 c37+70 C c17+84 c c40+70 c z35+14 z12 c c54+70 c26+70 c z35+56 c c12+42 c c18+84 z48 z32 z49+14 z65 z9 z49 z66 z25+16 z75 z34 z76 z67 z51 z42+14 z59 z68 z43 z78 z61 z79 z62 z m/z m/z

13 Protein Identification by Top Down MS/MS Me Me Ac Ac Ac Ac Ac N-AGRGK 5 GGK 8 GLGK 12 GGAK 16 RHRK 20 VLRDNIQGIT H4 Known Modification Ac Ac Ac Modification Detected $500 on Fusion or ICR

14 What is Imaging Mass Spectrometry? ImagingMassSpectrometryisatechnologythat combines advanced analytical techniques for the analysis of biological molecules with spatial fidelity. An effective approach for imaging biological specimens in this way utilizes Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI MS). Briefly, molecules of interest are embedded in an organic matrix compound that assists in the desorption and ionization of compounds on irradiation with a UV laser. The mass to charge ratio of the ions are measured using a mass spectrometer over an ordered array of ablated spots. Multiple analytes are measured simultaneously, capturing a representation or profile of the biological state of the molecules in that sample at a specific location on the tissue surface. Nature Reviews Cancer 10, (September 2010) doi: /nrc2917

15 $750/Sample Trends in Biochemical Science; Volume 29, Issue 3, March 2011, Pages

16 Problem Interference from Matrix Metabolites not Stable Solutions: High Mass Accuracy (FT ICR) Changing Matrix Derivatization Using Product Ions for Scanning $750/Sample on 15T ICR Biology 2015, 4(1),