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N-linked Glycan Characterization and Profiling: Combining the Power of Accurate Mass, Reference Glucose Units, and UNIFI Software for Confident Glycan Assignments

Waters
27 Mar 2014

This application note details a new workflow for a glycan assay, using FLR with mass confirmation, available in the latest version of the Biopharmaceutical Platform Solution with UNIFI. The practical use of this workflow for fluorescent labeled (2AB) N-linked released glycan characterization is illustrated using a biosimilar/innovator biotherapeutic comparability study.

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Biopharmaceutical Platform Solution with UNIFI

Waters

The Waters Biopharmaceutical Platform Solution with UNIFI® integrates robust UPLC®/MS characterization technology with the UNIFI Scientific Information System for applications in bioseparations, intact protein mass analysis, peptide mapping and released glycan analysis.This platform unites all aspects of biotherapeutic analyses and workflows, enabling organizations to apply high resolution analytics across the development process continuum. The platform captures complex mass spectrometry and chromatography data, and enables laboratories to comprehensively and routinely acquire, process, and share LC and MS characterization information throughout a biotherapeutic’s lifecycle. Results can be shared from discovery to development to GxP quality control.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.LC-MSLC-MS (liquid chromatography-mass spectrometry) systems and equipment are used for separation and quantitative analysis of complex mixtures, combining liquid chromatography and mass spectrometry. Quantify proteins, contaminants, pesticides or screen for drug metabolites with a high level of sensitivity. LC-MS systems and equipment include reverse phase, normal phase and specialized columns integrated with various MS detectors such as time-of-flight (TOF), quadrupole, orbitrap or ion trap mass analyzers. LC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzer give greater sensitivity and resolving power to your analysis. Find the best LC-MS equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.GlycobiologyGlycobiology encompasses research into the biological functions, synthesis and structures of glycans, including glycoproteins, glycolipids, proteoglycans and free oligosaccharides. Mass spectrometers and their software are commonly used in the study of glycobiology, along with structure predicting tools, chromatography and NMR.Data AnalysisData analysis hardware and software is available to make data processing straight-forward yet powerful. Data software can be used for math and stats, technical graphing and image analysis. In addition, software is available for specific data analysis of electrophoresis, densitometry, ELISA and DNA sequencing.Chem / BioinformaticsCheminformatics and bioinformatics are computational techniques used in chemistry and biology, respectively, for data acquisition, processing and storage. Cheminformatics focuses on compound information, whereas bioinformatics is mainly applied to analysis and modeling of genomics, genetic and sequencing information. Hardware and software is available for data acquisition, analysis, management and storage.BiosimilarsBiosimilars are very similar versions of approved and authorized medicines. Clinically inactive components will have minor differences, but there are no significant differences between the biosimilar and the original product in terms of potency, purity and safety.BiotherapeuticsBiotherapeutics are proteins and other compounds (such as nucleic acids) produced by living organisms that have uses as therapeutics or in <i>in vivo</i> diagnostics. The most well-known example of a biotherapeutic product, and the first to be approved for therapeutic use, was recombinant human insulin.Glycans

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