BioHPLC Column Selection Guide Cover - Agilent Technologies

ion-exchange Chromatography
4 www.agilent.com/chem/biohplc
iOn-eXCHanGe CHROMaTOGRaPHY
Purify proteins and other charged molecules
Ion-exchange chromatography (IEX) is a highly sensitive technique that allows you to separate ions
and polar molecules based on their charge. Like SEC, IEX can be used to separate proteins in their
native state.
Applying IEX to charge variant analysis
During production and purification, antibodies can exhibit changes in charge heterogeneity as a result
of amino acid substitutions, glycosylation, phosphorylation, and other post-translational or chemical
modifications. Because these changes can impact stability and activity – or cause immunologically
adverse reactions – the analysis of charge heterogeneity in monoclonal antibody (MAb) preparations
is critical to biopharmaceuticals.
In protein analysis, charge variations at a given pH indicate a change in the primary molecular
structure – resulting in additional forms of the protein in question. These are called isoforms
(or charge variants), and can be resolved by IEX chromatography. IEX is also useful as a
preparative technique.
The pages that follow describe Agilent’s family of weak and strong ion-exchangers – both anionic
and cationic.
• Agilent non-porous Bio IEX columns are designed for high-resolution, high-efficiency,
and high-recovery separations.
• Agilent Bio MAb columns are optimized for separating charge isoforms of monoclonal
antibodies.
• Agilent porous IEX columns (PL-SAX and PL-SCX) are chemically stable, and are available
in two pore sizes – allowing you to separate peptides, oligonucleotides, and very large proteins.
• Bio-Monolith IEX columns are uniquely suited to separating antibodies, viruses, and DNA.