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Simultaneous Determination of Molecular Size, Concentration, and Impurity Composition of Biotherapeutics with SEC and the Biopharmaceutical Platform Solution with UNIFI

Waters
27 Jul 2014

The objective of this application note is to demonstrate the ability to determine molecular weight and amount of the constituents of an antibody sample using UNIFI informatics. A purified antibody from human serum was used as a model protein to test the application.

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ACQUITY UPLC BEH Columns

Waters

In 2004, Waters revolutionized liquid chromatography with the introduction of the ACQUITY UPLC System. At the heart of the system was Waters second generation hybrid, the Ethylene Bridged Hybrid (BEH) particle. In 2005, BEH Technology preparative particle sizes (2.5, 3.5, 5 and 10 μm) with the introduction of XBridge between HPLC and UPLC Technology platforms. BEH Technology particles offer: Excellent peak shape and efficiency for basic analytes A rational array of chromatographic selectivity Improvements in chemical stability at mobile phase extremes, particularly at elevated pH

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Biopharmaceutical Platform Solution with UNIFI

Waters

The Waters Biopharmaceutical Platform Solution with UNIFI® integrates robust UPLC®/MS characterization technology with the UNIFI Scientific Information System for applications in bioseparations, intact protein mass analysis, peptide mapping and released glycan analysis.This platform unites all aspects of biotherapeutic analyses and workflows, enabling organizations to apply high resolution analytics across the development process continuum. The platform captures complex mass spectrometry and chromatography data, and enables laboratories to comprehensively and routinely acquire, process, and share LC and MS characterization information throughout a biotherapeutic’s lifecycle. Results can be shared from discovery to development to GxP quality control.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.UV-Visible SpectroscopyUltraviolet-visible (UV-Vis) spectrophotometers are used to measure the interaction of UV and visible light with a sample, including transmission, reflectance & absorbance. The two major instrument classes are single-beam or double-beam spectrophotometers. More specialized equipment includes colorimeters, spectroradiometers and refractometers. Portable and microvolume spectrophotometers are also available. For the modular spectroscopy lab, explore a range of light sources for combination with a spectrograph/spectrometer and optics. Find the best UV-Vis spectroscopy products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.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.Chromatography Method DevelopmentChromatography method development is the process of designing and optimizing chromatographic techniques to separate and analyze complex mixtures. This includes selecting appropriate stationary and mobile phases, determining flow rates, and optimizing detection methods. Successful method development is essential for high-quality results in fields like pharmaceuticals, food safety, and environmental analysis. Explore chromatography method development tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly 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.ManufacturingManufacturing is the production of merchandise for use or sale using labor and machines, tools, chemical and biological processing, or formulation.

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