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Development of Ultra-Fast pH-Gradient Ion Exchange Chromatography for the Separation of Monoclonal Antibody Charge Variants

18 May 2015

The pre-formulated buffers for pH gradient, introduced by Thermo Fisher Scientific, have greatly simplified the development of ion exchange chromatography (IEX) of monoclonal antibodies (mAbs). This application note shows how to push the throughput of pH gradient IEX even further. To achieve this, a MAbPac SCX-10 RS column, 5 μm, 2.1 × 50 mm was operated on a new Thermo Scientific™ Vanquish™ UHPLC system. The Vanquish UHPLC is a fully biocompatible system, suitable for the analysis of intact proteins.

Thermo Scientific™ MAbPac™ SCX-10 Monoclonal Antibody Analysis Column

Thermo Fisher Scientific

MAbPac™ SCX-10 Columns for the High-Resolution Separation and Analysis of Monoclonal Antibody VariantsThe Thermo Scientific™ MAbPac™ SCX-10 column line has been expanded to include 3 and 5 μm particle sizes for fast, high-resolution analysis as well as rapid separation (RS) columns for bio-UHPLC separations. These smaller-particle-size columns provide the same high resolution as the 10 μm columns, but with significantly faster analysis time. In addition, salt or pH gradients can be used to provide fast high-resolution, high-throughput analysis of monoclonal antibody (MAb) variant samples. The MAbPac™ SCX-10 columns are strong cation-exchange columns designed specifically for the high-resolution, high-efficiency analysis of MAbs and associated variants. The unique nonporous pellicular resin provides exceptionally high resolving power, permitting the separation of MAb variants that differ by as little as one charged residue. Hydrophobic interactions with the resin are eliminated for very efficient peaks.MAbPac™ SCX-10 Monoclonal Antibody Analysis Column Features: Unequalled high-resolution separations Exceptionally high efficiency Highest resolution of monoclonal antibody variants Unmatched column-to-column and lot-to-lot reproducibility Hydrophobic interactions eliminated Ideal for stability testing Column Details: Chemistry: Sulfonic Particle Size: 3, 5, 10 μm Length: 50, 150, 250 mm ID: 2.0, 4.0, 4.6 mm

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Development of Ultra-Fast pH-Gradient Ion Exchange Chromatography for the Separation of Monoclonal Antibody Charge Variants