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Characterization of the N-glycosylation Pattern of Antibodies by ESI - and MALDI Mass Spectrometry

21 Apr 2013

This application note describes the in-depth characterization of N-glycosylation pattern in various antibody samples using two complementary mass spectrometric strategies based on both, top-down ESI-UHR-TOF and bottom-up LC-MALDI-TOF/TOF performed on the Bruker maXis™ and ultrafleXtreme™ platform, respectively.

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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.Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. AntibodiesAntibodies are used in techniques such as confocal and fluorescence microscopy, flow cytometry, ELISA, ELISPOT, immunohistochemistry, western blotting and immunopreciptation. Select specific antigen reactivity, high specific affinity, low non-specific binding, monoclonal or polyclonal, primary or secondary antibodies and associated conjugates such as an enzyme or dye for visualization.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.MALDI-TOF MSMALDI-TOF mass spectrometers (matrix-assisted laser desorption/ionization time-of-flight) are used for the mass analysis of large molecules, including proteins, DNA, polymers and other macromolecules due to its relatively soft ionization, reducing fragmentation whilst allowing fast data acquisition. Tandem mass analyzers such as TOF/TOF systems can be used to increase resolution and sensitivity of your analysis. Find the best MALDI-TOF products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.BiopharmaceuticalsBiopharmaceuticals are proteins and other compounds (such as nucleic acids) produced by living organisms that have uses as therapeutics or for in vivo diagnostics. The most well known example of a biopharmaceutical product, and the first to be approved for therapeutic use, was recombinant human insulin.GlycosylationGlycoproteins
Characterization of the N-glycosylation Pattern of Antibodies by ESI - and MALDI Mass Spectrometry