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Reliable Results in Peptide Mapping Using the Vanquish Flex UHPLC System

Thermo Fisher Scientific
17 Nov 2015

Peptide mapping is one of the routine methods for biotherapeutics characterization. This technique, combined with mass spectrometry, is utilized in research environments for the determination of the primary sequence of an antibody or the identification of post-translational modifications. UV is often used as a unique detection tool, for instance in stability studies. The data interpretation of these experiments is based on retention times as qualitative and peak area as quantitative information. In this application note, the separation of peptides obtained from a bovine serum albumin (BSA) digest is demonstrated. The retention time and peak area precisions are evaluated for repeated injections.

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Thermo Scientific™ Vanquish™ Flex Quaternary UHPLC System

Thermo Fisher Scientific

The Thermo Scientific™ Vanquish™ Flex Binary UHPLC system is biocompatible with a state-of-the-art binary high-pressure pump and state-of-the-art binary solvent blending. This ultra high performance liquid chromatography system shares all Vanquish values, such as a design focused on uptime, robustness and reliability. Multiple detection options as well as Thermo Scientific™ Viper™ fingertight connections throughout the system will give you the performance you need with the ease of use you want. Our SmartFlow™ pumping technology with automatic compensation for changing eluent compressibility ensures excellent flow and gradient precision, independent of eluent composition and backpressures. This gives you unrivaled retention time precision and highest data confidence, even for ternary and quaternary gradients, and for more freedom in method development and application switching. Detection capabilities include high sensitivity and linearity Diode Array Detection with LightPipe technology, selective and sensitive Fluorescence Detection and near-universal detection capabilities with the Vanquish Charged Aerosol Detector. Key Features: Integrated modularity design concept to offer uncompromising UHPLC State-of-the-art quaternary solvent blending 1000 bar of pump pressure and flow rates up to 8 mL/min Industry-leading flow and gradient precision Multiple detection options including DAD, FLD and CAD Relevant Applications: Separations of Intact mAbs Analysis of labeled glycans Ternary gradient impurity profiling Efficient, reliable peptide mapping

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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.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.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.Peptide MappingPost-Translational ModificationsTherapeutic AntibodiesTherapeutic antibodies are antibodies engineered for use in treating diseases. They can be designed to target specific antigens, blocking harmful interactions or modulating immune responses. Therapeutic antibodies are critical in treating conditions like cancer, autoimmune disorders, and infectious diseases. Explore the best therapeutic antibody products in our peer-reviewed product directory; compare options, check reviews, and get pricing directly from manufacturers.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.Bovine Serum Albumin (BSA)

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Reliable Results in Peptide Mapping Using the Vanquish Flex UHPLC System