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Using Longer Aeris™ PEPTIDE Core-Shell HPLC/UHPLC Columns for Improved Peptide Mapping

Phenomenex Inc
11 Nov 2014

When one looks at peptide mapping applications, performance requirements are significantly different versus intact protein separations, as increased retention and selectivity are required to separate the large number of peptides generated in peptide mapping applications. Because increased resolution is a higher priority versus speed, a larger particle (3.6 μm) 100 Å HPLC/UHPLC column (Aeris PEPTIDE) was developed. It is specifically designed to improve separations of peptide and peptide mapping applications by allowing the use of longer columns at lower backpressures. In this application note, the increased resolution that the longer Aeris PEPTIDE 3.6 μm XB-C18 provides will be demonstrated.

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Aeris Core-Shell LC Columns for Proteins and Peptides

Phenomenex Inc

  Core-Shell Particles Precision Engineered for Protein and Peptide Separations.   Core-shell particle technology provides striking increases in peak capacity and resolution at lower backpressure, giving chromatographers the ability to achieve ultra-high performance on ANY system, HPLC or UHPLC. A uniform porous silica layer is grown around a solid, spherical silica core, providing effective retention and selectivity with improved resolution, speed, and recovery. Next, optimizing the pore size and shell thickness for intact proteins or smaller peptide fragments provides well-defined depth penetration of biomolecules leading to maximum separation power. Column Specifications: Phase: PEPTIDE XB-C18, WIDEPORE C4, WIDEPORE XB-C18, WIDEPORE XB-C8 Particle Size: 1.7, 2.6, 3.6, 5.0 µm Pore Size: 200, 100 Å Length: 50, 100, 150, 250 mm ID: 2.1, 3.0, 4.6, 10.0, 21.2 PEPTIDE XB-C18 Synthetic peptide impurity analysis Peptide mapping Identifying protein modifications - Glycosylation - Substitution - Truncation Analyzing post-translational modifications - Deamidation - Oxidation - Deletions WIDEPORE XB-C18 Proteins Hydrophilic proteins PEGylated proteins High temperature separations Alternative selectivity for peptide mapping WIDEPORE XB-C8 Large proteins Moderately hydrophobic proteins Monoclonal antibodies Glycosylated proteins High temperature separations WIDEPORE C4 Very large proteins Very hydrophobic proteins Membrane proteins Least retentive

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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.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.Peptide MappingProtein 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.

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Using Longer Aeris™ PEPTIDE Core-Shell HPLC/UHPLC Columns for Improved Peptide Mapping