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Advances in Size-Exclusion Chromatography for the Analysis of Small Proteins and Peptides: Evaluation of Calibration Curves for Molecular Weight Estimation

Waters
9 Apr 2013

Size exclusion chromatography has been the preferred method for the analyses of biomolecules based on size. The application note demonstrates the impact of a 125 Å pore sub-2 µm packing material on the separation and resolution of small proteins and peptides. The impact of both physical and chemical properties of SEC packing materials on SEC calibration curves used for molecular weight estimation is also demonstrated.

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ACQUITY UPLC H-Class PLUS Bio System

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The ACQUITY UPLC H-Class PLUS Bio System delivers the benefits of UPLC's resolution, sensitivity, and throughput in a system purpose-built for the analysis of proteins, peptides, nucleic acids, and glycans.

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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.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.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.TherapeuticsTherapeutics are treatments designed to alleviate or cure diseases. These include pharmaceuticals, biologics, and gene therapies, which work by targeting specific disease mechanisms. Advances in personalized medicine and biologics offer new hope for patients with conditions that were previously difficult to treat. Browse our peer-reviewed product directory to find therapeutic solutions for various diseases; compare products, check user reviews, and get pricing directly from manufacturers.

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