Waters Corporation · NYSE:WAT
Waters — Technical Presentation (2011)
24 slides from Waters's technical presentation, 2011
Slide 1: 2D UPLC Analysis of Proteins
2D UPLC Analysis of Proteins Patrick Boyce Pharmaceutical Marketing Team Manager Europe & India, Waters ©2011 Waters Corporation 1
Slide 2: Overview
Overview Technology Uses of 2D LC Application – mAb in cell culture media ©2011 Waters Corporation 2
Slide 3: ACQUITY UPLC Multi pump/Multi valve Solutions
ACQUITY UPLC Multi pump/Multi valve Solutions ACQUITY UPLC ACQUITY UPLC UPLC with Online System with 2D Automated SPE SPE Technology Technology System ©2011 Waters Corporation 3
Slide 4: ACQUITY Systems with 2D Technology
ACQUITY Systems with 2D Technology Do you need more sensitivity? Do you want to reduce the down time (cleaning) of your MS detector? Do you spend too much time with sample preparation? Would you like to improve the quality of your separation? The first ever ACQUITY UPLC System with 2D Tech
Slide 5: Customer Enabled 2D System
Customer Enabled 2D System ©2011 Waters Corporation 5
Slide 6: Purposefully Innovations to enable ACQUITY UPLC with 2D Technology
Purposefully Innovations to enable ACQUITY UPLC with 2D Technology ©2011 Waters Corporation 6
Slide 7: ACQUITY UPLC with 2D Technology
ACQUITY UPLC with 2D Technology ©2011 Waters Corporation 7
Slide 8: 3 types of applications
2D Chromatography With UPLC What Is This? 3 types of applications Online Sample Parallel Column Real 2D LC Preparation Regeneration Column 2 is being regenerating while column 1 is used for the analysis Increase sample throughput More flexibility, Faster ROI, Faster Decision Making, Less Waiting Tim
Slide 9: 50% Increase in Throughput
Business Benefits – Parallel Column Regeneration Contract Research Lab performing Metabolite Profiling looking to improve throughput Typical UPLC UPLC with 2D Technology Injection to Injection 12 Minutes 8 Minutes Cycle Time Samples Analyzed in 120 180 24 Hours 8 Minute Separation and 4 Minute Re-Eq
Slide 10: 3 types of applications
2D Chromatography With UPLC What Is This? 3 types of applications Parallel Column Real 2D LC Online Sample Preparation Regeneration Column 2 is being Trap & Elute regenerating while column 1 is used for the analysis Sample of interest is Impurities can be trapped Increase sample trapped and concentr
Slide 11: Increase Sample Loading and Sensitivity with Trap and Elute Configuration
Increase Sample Loading and Sensitivity with Trap and Elute Configuration MRM of 6 Channels ES+ 2.14 327 > 269.9 100 1.80e5 20 uL Injection Clozapine 1 ppb on 1D System % 0 Time 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 MRM of 6 Channels ES+ 100 2.73 327 > 269.9 4.28
Slide 12: 3 types of applications
2D Chromatography With UPLC What Is This? 3 types of applications Parallel Column Real 2D LC Online Sample Preparation Regeneration Column 2 is being Heart Cutting Trap & Elute regenerating while column 1 is used for the analysis 1/Sample passes through the first column Sample of interest is trapped
Slide 13: Typical LC Separation versus Separation Using 2D Heart Cutting
Typical LC Separation versus Separation Using 2D Heart Cutting Typical LC Separation ACQUITY UPLC with 2D Technology desired peptides phospholipids (co-eluting) interference desired peptides no phospholipids ©2011 Waters Corporation 13
Slide 14: Two Dimensional UPLC Applications
Two Dimensional UPLC Applications Application Note – Quantification and MS analysis of monoclonal antibodies in complex samples. Improve productivity by monitoring mass profiles and yields during clone selection and production scale-up from a single injection Poster ASMS 2012 – Lipid Profiling o
Slide 15: Rapid Quantification and MS Analysis of Monoclonal Antibodies in a Complex Samples
Rapid Quantification and MS Analysis of Monoclonal Antibodies in a Complex Samples ©2011 Waters Corporation 15
Slide 16: Challenge – Quantitative Analysis of Monoclonal Antibodies
Challenge – Quantitative Analysis of Monoclonal Antibodies Analysis of Monoclonal Antibodies in Cell Culture Media — Complex matrix Fraction Collection and Reinjection Challenges — Time consuming — Labor intensive — Degradation of antibodies or adsorption to collection vessel ©2011 Waters Corpor
Slide 17: 2D UPLC Quantitative Analysis of Monoclonal Antibodies
2D UPLC Quantitative Analysis of Monoclonal Antibodies Objective — Quickly and Effectively Analyze mAb titer and profile from cell culture media in a single injection Approach — 2D UPLC Heart Cut and Trap and Back-Flush Elution — ACQUITY UPLC H-Class Bio System with 2D Technology — TUV Detector
Slide 18: Experimental
Experimental Injection: 0.5-10 µL mAb cell culture media First dimension (2.1x30mm protein A affinity column) — Mobile phase A: 50mM phosphate pH 7.0, 150mM NaCl — Mobile phase B: 12mM HCl, pH 1.9, 150mM NaCl — Flow rate 1.0mL/min — Gradient 0-100% B over 1 minute Second dimension (2.1mmx5mm M
Slide 19: First Dimension: Isolation of Monoclonal Antibodies on Protein A Column
First Dimension: Isolation of Monoclonal Antibodies on Protein A Column Peak Collection Window mAb Overlay of 12 injections Peak Area 0.078 % RSD Flow Through Blank Injection 0 1.0 2.5 Minutes ©2011 Waters Corporation 19
Slide 20: Response Linearity with Mass Load
Response Linearity with Mass Load ©2011 Waters Corporation 20
Slide 21: Sample Raw MS from desalted Monoclonal Antibodies
Sample Raw MS from desalted Monoclonal Antibodies ©2011 Waters Corporation 21
Slide 22: Deconvoluted MS
Deconvoluted MS G0F/G1F G1F/G1F G0F/G0F G1F/G2F G0/G0F G2F/G2F ©2011 Waters Corporation 22
Slide 23: Benefits of 2D UPLC Monoclonal Antibody Analysis
Benefits of 2D UPLC Monoclonal Antibody Analysis Reduces Sample Handling — Reduce Possibility of Sample Loss — Reduce Possibility of Sample Degradation Maximize MS Response Improve separation Rapid Isolation and Determination of mAb concentration and profile from cell culture media ©2011 Wat
Slide 24: Thank You!
Thank You! ©2011 Waters Corporation 24