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Separation of Intact Monoclonal Antibody Sialylation Isoforms by pH Gradient Ion-Exchange Chromatography

28 Aug 2014

This application note demonstrates high-resolution separation of intact protein sialylation isoforms using the Thermo Scientific™ ProPac™ SCX-10 column on a Thermo Scientific™ Dionex™ UltiMate™ 3000 Dual BioRS LC System with a salt or pH gradient.

Thermo Scientific™ ProPac™ SCX and WCX Protein Column

Thermo Fisher Scientific

Strong and Weak Cation-Exchange Columns for High-Resolution Separation of Proteins and MAbsThe Thermo Scientific ProPac™ SCX column line has expanded to include the next generation ProPac SCX-20 column. This column provides unequalled high resolution of protein variants.ProPac SCX-10 and WCX-10 strong and weak cation-exchange columns are based on a nonporous core particle that results in high-efficiency peaks and high-resolution separations of protein variants. A hydrophilic layer prevents unwanted secondary interactions. A grafted cation-exchange surface provides pH-based selectivity control and fast mass transfer for high-efficiency separation and moderate capacity.ProPac® SCX and WCX Protein Column Features: Characterization and quality control assessment of monoclonal antibodies and other proteins Unequalled resolution High-resolution separations with excellent sample recovery Unmatched column-to-column and lot-to-lot reproducibility Useful for characterization of related protein variants (e.g., deamidation and MAb lysine truncation variants) Highly-efficient peaks with excellent sample recovery Available in different dimensions and formats for flexibility in method development Column Details: Length: 50, 100, 150, 250 mm ID: 2.0, 4.0, 9.0, 22 mm

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Ion ChromatographyIon chromatography, also known as ion exchange chromatography, is a high-performance liquid chromatographic technique used for the separation and identification of ions or polar molecules in a sample, including proteins, nucleotides and amino acids. Equipment includes ion exchange columns, ion exclusion columns, ion chromatography systems, pumps, and detectors. Find the best ion chromatography equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.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.GlycosylationProtein BiologyThe analysis of protein expression, identity and function is vital for many areas of life science research and drug discovery. Some of the most commonly used techniques in protein analysis include Western blotting, electrophoresis and mass spectrometry.Monoclonal AntibodiesMonoclonal antibodies (mAbs) are lab-made molecules that can bind to specific antigens on cells, making them highly effective in treating cancers, autoimmune diseases, and infections. These targeted therapies are transforming the treatment landscape for many conditions. Browse our peer-reviewed product directory to find monoclonal antibodies for research and clinical applications; compare products, check reviews, and get pricing directly from manufacturers.