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Precise Characterization of Intact Monoclonal Antibodies

Agilent Technologies
21 Aug 2019

Monoclonal antibodies (mAbs) are a very important class of biopharmaceutical molecules. As a protein drug, thorough characterization of the mAb is required in each of the manufacturing steps. Intact mAb analysis offers rapid assessment on determining the accurate molecular weight of an mAb product and its degree of heterogeneity, such as post-translational modifications (PTMs), antibody-drug conjugate (ADC), mAb sequence variations, or degradation products. Quadrupole Time-of-flight (Q-TOF) LC/MS systems are often used to analyze intact proteins or antibodies due to excellent resolution at the high mass range. This application note describes a seamless workflow using the Agilent 1290 Infinity II UHPLC system, 6545XT AdvanceBio LC/Q-TOF, and automatic data processing with Agilent MassHunter BioConfirm software to analyze a variety of mAb products.

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Agilent 1290 Infinity II LC System

Agilent Technologies

The 1290 Infinity II LC System is an ultra-high performance LC that has market-leading sample capacity and is designed for the lowest dispersion for the most challenging ultra-HPLC experiments. The 1290 Infinity II LC system is designed to be both a HPLC and high-end UHPLC system in a single flow path, giving you robust and excellent performance no matter what application you are running while seamlessly integrating into your existing laboratory environment.

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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.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.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.

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