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Full Validation of Therapeutic Antibody Sequences by Middle-Up Mass Measurements and Middle-Down Protein Sequencing
Full Validation of Therapeutic Antibody Sequences by Middle-Up Mass Measurements and Middle-Down Protein Sequencing
12 Oct 2016This application notes reports a novel middle-up LC-ESI-MS subunit analysis and middle-down LC-MALDI in-source decay (ISD) mass spectrometry method applied to cetuximab, panitumumab and natalizumab selected as representative FDA and EMA approved monoclonal antibodies (mAbs). The regulatory agencies require comprehensive primary sequence validation both for innovator and biosimilar monoclonal antibodies (mAbs). Full sequence coverage (SC) is typically obtained by the combination of multiple LC-MS/MS datasets from various protease digests.
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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.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.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.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.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.Protein DetectionProtein detection techniques are essential for understanding protein expression, localization, and function. Methods such as western blotting, ELISA, and mass spectrometry are commonly used in research and diagnostics. Browse our peer-reviewed product directory to find the best tools for protein detection, compare products, check reviews, and get pricing directly from manufacturers.ASMSThe American Society for Mass Spectrometry (ASMS) is an organization dedicated to advancing the science and application of mass spectrometry. ASMS supports professionals through events, publications, and research initiatives, promoting innovation in areas like proteomics, metabolomics, and environmental analysis. Antibody ProductionAntibody production is essential for a wide range of applications in biochemistry and immunology. Ensuring high-quality monoclonal and polyclonal antibodies are produced is key. Explore antibody production tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Clinical Mass SpectrometryMass spectrometry is emerging as a superior method of analysis over the longstanding, gold standard of immunoassay. Clinical mass spectrometry systems are increasingly accessible to medical laboratories and are capable of providing results with more speed and less expense than traditional methods. From simple mass spectrometers to IVDR class A compliant, highly sophisticated systems performing liquid chromatography tandem mass spectrometry (LC/MS/MS), find the latest product news, reviews and resources to make a purchasing decision for your laboratory.