EC50 Cartridges

The Ludger EC50 cartridges have been developed as a lower cost replacement for the LudgerClean EB10 cartridges while having equivalent efficiencies and reproducibility.

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The cartridges contain an electron interaction resin (EIR). This unique material contains very flat regions of pi-orbital electrons that interact with hydrophilic organic compounds including glycans.

The resin can capture a wide range of glycans from complex mixtures while salts, proteins, and certain detergents pass through. The glycans are then eluted from the cartridge using a simple solvent system.

2-AB Labeling Efficiency: Use of EC50 Cartridges vs No Clean-up

UHPLC HILIC profile of 2-AB labeled human IgG standards in PBS – 11 replicates;
Black: 2-AB labeling after EC-50 clean-up. Blue: 2-AB labeling with no clean-up performed.

Average absolute area of the human IgG peaks (11 replicates) with EC50 clean-up (black)
and with no clean-up (blue) followed by 2-AB labeling/T1 clean-up

Ludger EC50 Cartridge Reproducibility

UHPLC HILIC profile of 2-AB labeled human IgG – 11 replicates; post EC-50 clean-up of CLIBN-IGG-5U in PBS

EC50 and EB10 Equivalence

Average relative % of human IgG standard peaks post EB10 (8 replicates)
and post EC50 (11 replicates) clean-up comparison

Average relative area % of human IgG standard peaks post EB10 (8 replicates) and post EC50 (11 replicates)
followed by 2-AB labeling and post-labeling cleanup (T1 cartridges)

Binding Capacity Each EC50 cartridge can typically bind up to 50 μg of O- or N-linked glycans.

Number of Samples Ludger EC50 cartridges are designed for single use only.

Suitable Samples A wide range of glycans can be purified. These include N-linked and O-linked type oligosaccharides, tri-saccharides and larger structures.

The cartridges are not suitable either for monosaccharides or disaccharides which are generally bound too weakly for efficient purification or for large linear poly-sialylated glycans which can be bound very tightly to the resin.

Glycan samples must be applied to the cartridges in solutions that are substantially aqueous.

Structural Integrity No detectable (< 2 mole per cent) loss of sialic acid, fucose, sulfate, or phosphate.

Binding efficiency >95% for most glycans

Binding Selectivity Essentially stoichiometric binding and elution for most complex glycan mixtures.

Storage Store at room temperature in the dark. Protect from sources of heat, light, and moisture. The cartridges are stable for at least two years as supplied.

Shipping The product can be shipped at ambient temperature.

Handling Ensure that any glass, plastic ware or solvents used are free of glycosidases and environmental carbohydrates. Use powder-free gloves for all sample handling procedures and avoid contamination with environmental carbohydrate.

Safety Please read the Safety Data Sheet (SDS) for all chemicals used.

All processes involving hazardous reagents should be performed using appropriate personal safety protection – eyeglasses, chemically resistant gloves (e.g. nitrile), etc. – and where appropriate in a laboratory fume hood.