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Native SEC/MS analysis of mAbs on an entirely biocompatible flow path

23 Mar 2023

Size exclusion chromatography (SEC) of recombinant therapeutic proteins, such as monoclonal antibodies (mAbs), is an essential analytical tool to detect the presence of size variants, which may have adverse effects on safety and efficacy. SEC is commonly carried out using nonvolatile phosphate buffers, which makes the separation incompatible with mass spectrometry (MS). For this reason, a desalting step (online or offline) is required for peak identification. In this application note, learn more about the potential of the combination of a polyether ether ketone (PEEK)-lined Agilent AdvanceBio SEC column with the Agilent 1290 Infinity II Bio LC System for native SEC/MS analysis of mAbs.

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Size Exclusion ChromatographySize exclusion chromatography (SEC) is a liquid chromatographic technique used to separate and identify molecules based on their size, including polymers, proteins, nanoparticles & nucleic acid. SEC is further subdivided into gel filtration chromatography (GFC) and gel permeation chromatography (GPC) when an aqueous or organic mobile phase is used respectively. HPLC-based SEC systems and columns are available for analytical separation alongside SEC spin columns or filters to prep samples for many protocols. Find the best size exclusion 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.ImmunologyImmunological techniques measure and characterize immune responses. Immunology kits and analysis systems often use techniques such as ELISA, radioimmunoassay (RIA) and immunodiffusion assays, Immunohistochemistry, and flow cytometry. Immunologists use equipment such as flow Cytometers, plate readers, plate washers and fluorescent microscopes.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. 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.SECSize exclusion chromatography (SEC) separates molecules based on size, ideal for protein purification, polymer analysis, and biopharma applications. SEC ensures precise characterization of biomolecules for research and industry. Explore peer-reviewed SEC products and compare options for your lab.
Native SEC/MS analysis of mAbs on an entirely biocompatible flow path