ResourceSeparations

Analysis of Proteins by Size-Exclusion Chromatography Coupled with Mass Spectrometry Under Non-Denaturing Conditions

20 Oct 2013

This application note describes SEC-MS under non-denaturing conditions. While this application has been evaluated for SE-HPLC, UPLC Technology in combination with sub-2-μm SEC column packing and a time-of-flight mass spectrometer (Xevo G2 Q-Tof) allows for direct analysis with improved resolution and sensitivity. By combining MS conditions with a Xevo G2 Q-Tof, SE-UPLC-MS analysis can be used to assist in the identification of unknown components in a biotherapeutic.

ACQUITY UPLC H-Class PLUS Bio System

Waters

The ACQUITY UPLC H-Class PLUS Bio System delivers the benefits of UPLC's resolution, sensitivity, and throughput in a system purpose-built for the analysis of proteins, peptides, nucleic acids, and glycans.

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BEH (Ethylene Bridged Hybrid) Particles for ACQUITY UPLC

Waters

The 1.7 µm Ethylene Bridged Hybrid [BEH] particle is one of the key enablers behind UPLC® technology.It is available in three different pore sizes [130Å, 200Å and 300Å] and several bonded phases for reversed-phase and hydrophilic interaction chromatography and is applicable from small molecule to large biopharmaceutical analysis. Due to the intrinsic chemical stability of hybrid particle technology, a wider usable pH range [pH 1-12] can be employed, enabling a versatile and robust separation technology for method development. BEH particle technology is available in HPLC particle sizes [2.5, 3.5, 5 and 10 µm] in the XBridge® family of HPLC columns, enabling seamless transfer between HPLC and UPLC technology platforms.

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MassLynx MS Software

Waters

Acquire, Analyze, Manage, and Share Mass Spectrometry Information MassLynx™ Software increases the speed at which you can convert your sample data into valuable knowledge. It provides you with the fundamental platform to acquire, analyze, manage, and share your mass spectrometry information. MassLynx intelligently controls any Waters mass spectrometry system, from sample and solvent management components to mass spectrometer and auxiliary detectors. MassLynx Software may acquire nominal mass, exact mass, MS/MS and exact mass MS/MS data. MassLynx’s Sample List is the core of the system. It maintains and consolidates the data on all of your samples. You also initiate any activities related to your sample from the Sample List. This "sample centric" approach simplifies the interaction with your LC/MS or GC/MS system, acquired data and processed results. Process Application-specific Data MassLynx Software features general purpose and specialized Application Managers that provide information for your specific MS analyses and data. Two come standard with MassLynx: QuanLynx™ for automated quantification included as standard with MassLynx OpenLynx™ for qualitative screening and identification Optional Application Managers can perform: Targeted quantitative analysis Mass-directed purification Metabolite identification Deconvolution of complex chromatograms Metabonomics & metabolomics Protein identification and protein characterization

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