Separation Science Built for Biopharma
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1 Separation Science Built for Biopharma Pharma & Biopharma Tours 2016 The world leader in serving science
2 Overview 1. Evolution of UHPLC built for biopharma Meet the family 2. What makes Vanquish different? Key features and benefits 3. Biopharma workflows 5 fundamental workflows 2
3 Evolution of UHPLC Built for Biopharma 3
4 Specificity of Action Why Focus on Biopharmaceuticals? Small molecule Chemotherapeutic agent Small protein Antibody Antibody-Drug Conjugate Analytical Complexity 4
5 We Listened more sensitivity easier method development more resolution more productivity easier interaction & operation a system that was Built for Biopharma more robust & reliable more sample capacity fresh & exciting instruments faster service more access to experts 5
6 Meet the Family Thermo Scientific UltiMate 3000 BioRS UHPLC System Established biocompatible UHPLC Thermo Scientific Vanquish Flex UHPLC System New generation biocompatible UHPLC (quaternary pump, 1000 bar) Thermo Scientific Vanquish UHPLC System New generation biocompatible high-end UHPLC system (binary pump, 1500 bar) 6
7 What Makes Vanquish Different? 7
8 Innovation in Every Aspect Dynamic piston seal DE A1 Metering device dual drive DE A1 Sample pre-compression DE B4 Vial aeration DE B3 Double conical needle seat DE A1 Static flange seal DE A1 Light pipe flow cell core DE B3 Light fibre to fluid channel connection DE A1 Detector cooling control DE A1 Alignment of optical channel DE B3 Fitting - Nanoviper DE B3 Fitting - Viper DE B4 Alignment of rotor and stator DE B4 Rotor position measurement DE A1 26 filed Blank holder patent mechanics families / granted patents in 19 patent Spherical design of the rotor DE B3 DE B4 families Valve bearing system DE A1 Method for compensating thermal effects DE A1 Autofrettage at pump head DE B4 Piston coupling DE B4 Inline piston position measurement DE A1 High pressure check valve DE B4 High pressure pump head connection DE B4 Static sealing system for mixers DE A1 Radial mixer DE B3 Fitting Viper 3 DE B3 8
9 What Makes Vanquish Different? Tool-free fluidics Compact footprint Intuitive fluidic layout Easy switching of modules/ maintenance 9
10 What Makes Vanquish Different? Tool-free fluidics Compact footprint Intuitive fluidic layout Simplicity Easy switching of modules/ maintenance 10
11 What Makes Vanquish Different? Tool-free fluidics Compact footprint Intuitive fluidic layout Simplicity Easy switching of modules/ maintenance Precise flow rate and gradient delivery Ultra low pump pressure ripple Thermo Scientific LightPipe Flow Cell Accurate temperature control Intelligent sample precompression 11
12 What Makes Vanquish Different? Tool-free fluidics Compact footprint Intuitive fluidic layout Simplicity Easy switching of modules/ maintenance Precise flow rate and gradient delivery Ultra low pump pressure ripple Performance Thermo Scientific LightPipe Flow Cell Accurate temperature control Intelligent sample precompression 12
13 What Makes Vanquish Different? Tool-free fluidics Compact footprint Intuitive fluidic layout Simplicity Easy switching of modules/ maintenance Precise flow rate and gradient delivery Ultra low pump pressure ripple Performance Thermo Scientific LightPipe Flow Cell Accurate temperature control Intelligent sample precompression Customizable gradient delay volume Multiple heating modes Large sample capacity Broad flow rate and temperature range 4 x detectors and simple MS integration Supports long column capacity & all major chemistries 13
14 What Makes Vanquish Different? Tool-free fluidics Compact footprint Intuitive fluidic layout Simplicity Easy switching of modules/ maintenance Precise flow rate and gradient delivery Ultra low pump pressure ripple Performance Thermo Scientific LightPipe Flow Cell Accurate temperature control Intelligent sample precompression Customizable gradient delay volume Multiple heating modes Large sample capacity Versatility Broad flow rate and temperature range 4 x detectors and simple MS integration Supports long column capacity & all major chemistries 14
15 What Makes Vanquish Different? Tool-free fluidics Compact footprint Intuitive fluidic layout Simplicity Easy switching of modules/ maintenance Precise flow rate and gradient delivery Ultra low pump pressure ripple Performance Thermo Scientific LightPipe Flow Cell Accurate temperature control Intelligent sample precompression Customizable gradient delay volume Multiple heating modes Large sample capacity Versatility Broad flow rate and temperature range 4 x detectors and simple MS integration Supports long column capacity & all major chemistries Barcode reader Dry-air sample cooling Precision Thermo Scientific Dionex Chromeleon Chromatography Data System (CDS) Software 15
16 What Makes Vanquish Different? Tool-free fluidics Compact footprint Intuitive fluidic layout Simplicity Easy switching of modules/ maintenance Customizable gradient delay volume Multiple heating modes Large sample capacity Versatility Broad flow rate and temperature range 4 x detectors and simple MS integration Supports long column capacity & all major chemistries Biopharma Precise flow rate and gradient delivery Ultra low pump pressure ripple Performance Thermo Scientific LightPipe Flow Cell Accurate temperature control Intelligent sample precompression Barcode reader Dry-air sample cooling Precision Thermo Scientific Dionex Chromeleon Chromatography Data System (CDS) Software Confidence 16
17 Integration in Your Laboratory AppsLab application library Columns, Prep and Thermo Scientific Viper finger-tight connections Thermo Scientific Dionex Chromeleon Chromatography Data System (CDS) Software Fully integrated mass spectrometry solutions Seamless method transfer 17
18 Chromeleon Increase productivity Industry-leading Operational Simplicity and throughput with eworkflows Faster data processing, visualization, and reporting with dynami updating and Intelligent Functionality built-in Control all your instrumentation Thermo Scientific LC/IC/GC/ (quant) MS plus >300 third-party modules Customizable instrument epanels for consistent look and feel Achieve regulatory compliance Flexible to support research and compliant laboratories Comprehensive user management and esignatures Simplify administration Centralized or distributed data storage Scales from workstation to large global networks Connect your laboratory Easily interface with other laboratory software (e.g. LIMS) Download ready-to-run applications from AppsLab Library 18
19 Range of Detectors Thermo Scientific Vanquish Diode Array Detector (DAD) Thermo Scientific Vanquish Variable Wavelength Detector (VWD) Thermo Scientific Vanquish Fluoresence Detector (FLD) 4 modular detector options Can be used discretely or in series 1. DAD with LightPipe technology outstanding signal to noise performance and linearity 2. VWD supporting a wide operating range 3. FLD with high detection precision 4. CAD offers a sensitive, universal detection with a near uniform response Thermo Scientific Vanquish Charged Aerosol Detector (CAD) 19
20 Range of Detectors Thermo Scientific Vanquish Diode Array Detector (DAD) 4 modular detector options Can be used discretely or in series 1. DAD with LightPipe technology outstanding signal to noise performance and linearity Thermo Scientific Vanquish Variable Wavelength Detector Sensitivity, linearity, (VWD) and the 2. VWD supporting a wide operating range flexibility to switch Thermo Scientific Vanquish Fluoresence Detector (FLD) 3. FLD with high detection precision 4. CAD offers a sensitive, universal detection with a near uniform response Thermo Scientific Vanquish Charged Aerosol Detector (CAD) 20
21 Fully Integrated MS Solutions Hardware Dedicated connection kit Dedicated connection kit connects the LC to the MS Thermo Scientific Orbitrap Tribid Lumos MS Thermo Scientific Q Exactive MS Series Thermo Scientific TSQ MS Series Thermo Scientific LTQ MS Series The column compartment can be located at either side of the LC instrument for optimal flow paths Control and processing depending on user preference Thermo Scientific Xcalibur (with Thermo Scientific SII) Chromeleon CDS In both cases data can be exported for processing with other MS software packages Software Chromeleon SII for Xcalibur 21
22 Fully Integrated MS Solutions Hardware Dedicated connection kit Dedicated connection kit connects the LC to the MS Thermo Scientific Orbitrap Tribid Lumos MS Thermo Simple, Scientific stress-free on user preference MS Q Exactive MS Series connection Thermo Scientific TSQ MS Series Thermo Scientific LTQ MS Series The column compartment can be located at either side of the LC instrument for optimal flow paths Control and processing depending Thermo Scientific Xcalibur (with Thermo Scientific SII) Chromeleon CDS In both cases data can be exported for processing with other MS software packages Software Chromeleon SII for Xcalibur 22
23 Simple Method Transfer Gradient delay volume (GDV) adaption Adjustable metering device idle volume Mixer and Viper capillaries Delayed injection Match the thermostatting mode Forced and still air mode give different results Mimic thermal conditions used in existing systems UltiMate 3000 column compartments or that of other (U)HPLC systems 23
24 Simple Method Transfer Gradient delay volume (GDV) adaption Adjustable metering device idle volume Mixer and Viper capillaries Delayed injection Easy, fast method transfer & method development Match the thermostatting mode Forced and still air mode give different results Mimic thermal conditions used in existing systems UltiMate 3000 column compartments or that of other (U)HPLC systems 24
25 Biopharma Workflows 25
26 5 Fundamental Workflows Peptide mapping Aggregate analysis Charge variant analysis Intact protein analysis Glycan analysis Reversed Phase SEC IEX RP HILIC HIC HIC Mixed Mode Confirm sequence Check monomer vs Aggregates Check charge variation within antibody sample Check purity of the antibody Check glycan presence and structure UV and UV-MS UV (UV-MS) UV UV and UV-MS FLD, FLD-MS, CAD Analytical reproducibility Peak capacity High salt condition, biocompatible, low dispersion, reproducibility Biocompatible selectivity Biocompatible, gradient and thermostatting capabilities Gradient capabilities 26
27 1. Peptide Mapping mau Very high precision of both retention time and peak area Excellent separation in 30 minute gradient Active eluent pre-heating to avoid any thermal mismatch min 27
28 1. Peptide Mapping % RSD mau min 0.16 Very high precision of both 0.14 retention time and peak area Excellent separation in minute gradient 0.06 Active 0.04 eluent pre-heating to avoid any thermal 0.02 mismatch Peak # RSD 0.15% 0.05% 0.02% 28
29 1. Peptide Mapping % RSD mau Very high precision of both 0.14 retention time and peak area 0.12 Get the highest RT precision, 0.10 Excellent separation in minute gradient from discovery to QA/QC Active 0.04 eluent pre-heating to avoid any thermal 0.02 mismatch Peak # min RSD 0.15% 0.05% 0.02% 29
30 2. Aggregate Analysis MAbPac SEC x 150mm MAbPac HIC x 100mm Fast aggregate analysis Aggregate analysis with additional information 30
31 2. Aggregate Analysis MAbPac SEC x 150mm MAbPac HIC x 100mm Stop worrying about carry-over, discover the details Fast aggregate analysis Aggregate analysis with additional information 31
32 3. Charge Variant Analysis 180 mau min ph Gradients allow generic gradients for the separation of mabs with a range of isoelectic points Dedicated buffer kit allows easy method set-up Easy to optimize to increase sample through-put 32
33 3. Charge Variant Analysis 180 mau ph Gradients allow generic gradients for the separation of mabs with a range of isoelectic points Perform 160 rapid, reproducible ph 140 gradients min Dedicated buffer kit allows easy method set-up Easy to optimize to increase sample through-put 33
34 4. Intact Protein Analysis stressed reference Fast method for stability evaluation of intact antibodies Boosted separation performance through extended temperature range Highly efficient separation of intact antibodies and degradation products 34
35 4. Intact Protein Analysis stressed Fast method for stability evaluation of intact antibodies Boosted separation performance through extended temperature range Detect reference low-level degradants, fast Highly efficient separation of intact antibodies and degradation products 35
36 5. Glycan Analysis counts 2.0e8 Peak RSD % % % % % % Ternary gradient for mixed mode separation More than 60 peaks detected Average retention time RSD well below 0.1% min 36
37 5. Glycan Analysis counts 2.0e8 Peak RSD % % % Ternary gradient for mixed mode separation Easily develop methods for % % % maximum information More than 60 peaks detected Average retention time RSD well below 0.1% min 37
38 Summary Built to exacting standards, specifically for biopharma Simply, you can have the most versatile, high performance UHPLC that gives you more confidence in your results Unlock total workflow solutions for peptide mapping, glycans, intact protein analysis, charge variant profiling and aggregate analysis 38
39 Thank you 39
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