Improving Intact IgG Separations with Aeris™ WIDEPORE Core-Shell HPLC/UHPLC Columns
3 Nov 2014A widepore core-shell HPLC/UHPLC column (Aeris WIDEPORE) has been introduced that is specifically designed to improve protein separations. Rather than utilize a similar morphology of small molecule core-shell columns with larger pores, a completely different particle was developed that takes into account the slower diffusion of proteins into porous particles. When one looks at different intact protein separations, the resolution of different glycoforms of therapeutic IgG antibodies probably stands out as one of the more difficult due to the large size and structure of IgG. In this article improved separation of IgG is shown using Aeris coreshell HPLC/UHPLC columns.
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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