Using Longer Aeris™ PEPTIDE Core-Shell HPLC/UHPLC Columns for Improved Peptide Mapping
11 Nov 2014When 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 IncCore-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