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Advances in Size Exclusion Chromatography for the Analysis of Macromolecular Proteins

Waters
1 Apr 2013

Size exclusion chromatography has been the preferred method for the analysis of proteins based on size. This application note demonstrates how by combining 450 Å sub 3-µm packing materials with a low dispersion ACQUITY UPLC H-Class System, separations with improved resolution and high-throughput of SE-UPLC can be realized for macromolecular proteins and highly aggregated proteins with molecular weights of up to approximately 2000 kDa.

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BEH (Ethylene Bridged Hybrid) Particles for ACQUITY UPLC

Waters

The 1.7 µm Ethylene Bridged Hybrid [BEH] particle is one of the key enablers behind UPLC® technology.It is available in three different pore sizes [130Å, 200Å and 300Å] and several bonded phases for reversed-phase and hydrophilic interaction chromatography and is applicable from small molecule to large biopharmaceutical analysis. Due to the intrinsic chemical stability of hybrid particle technology, a wider usable pH range [pH 1-12] can be employed, enabling a versatile and robust separation technology for method development. BEH particle technology is available in HPLC particle sizes [2.5, 3.5, 5 and 10 µm] in the XBridge® family of HPLC columns, enabling seamless transfer between HPLC and UPLC technology platforms.

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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.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.AggregationProtein BiologyThe analysis of protein expression, identity and function is vital for many areas of life science research and drug discovery. Some of the most commonly used techniques in protein analysis include Western blotting, electrophoresis and mass spectrometry.Molecular Weight AnalysisMolecular weight analysis determines the size and structure of biomolecules, essential in protein characterization, polymer studies, and drug development. Techniques like mass spectrometry ensure precise results. Compare solutions for molecular weight analysis in our peer-reviewed product directory.PeptidesPeptides are short chains of amino acids that play key roles in biological processes, including signaling, immune responses, and enzyme regulation. Peptide research is critical in drug discovery, diagnostics, and vaccine development. Browse our peer-reviewed product directory to find the best peptides and peptide-related products, compare options, check reviews, and get pricing directly from manufacturers.

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Advances in Size Exclusion Chromatography for the Analysis of Macromolecular Proteins